Bergamo® II Series Multiphoton Microscopes


Bergamo® II Series Multiphoton Microscopes


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Sam Tesfai
Sam Tesfai
General Manager,
Thorlabs Imaging Systems

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Bergamo® II Series Multiphoton Microscopy Platform

Following the principle that the microscope should conform to the specimen, rather than the other way around, we created a completely modular multiphoton imaging platform that adapts to a wide range of experimental requirements. Our multiphoton imaging systems enable simultaneous readout and manipulation of neuron populations at higher speed, greater depth, and higher intensity than with traditional research techniques, such as using microelectrodes. As shown in the Options at a Glance section below and in the Five-Axis Movement of Rotating Bergamo Microscopes video to the lower right, we have developed a rotating body, which allows the microscope to rotate around the sample. We also offer a selection of rigid bodies, which feature an industry-leading 7.74" throat depth for a large three-dimensional working volume around the objective.

The features of Bergamo II listed in the Highlights, Features, and Modules tabs reflect our focus on developing cutting-edge capabilities without compromising usability. Detailed summaries of all the recommended applications for the Bergamo microscope can be found in the Applications tab. To get users started on designing their own microscope, we provide the Configurations tab, which lists a number of example configurations with key features and suggested applications. This platform can be easily modified or upgraded as experimental needs evolve; see the available add-ons and upgrades in the Retrofits tab. 

Bergamo imaging systems have been used to capture impressive sample images (see the Image Gallery tab) and have been featured in numerous publications (see the Publications tab). As shown in the Innovation through Collaboration video to the lower right, the Thorlabs Life Sciences division partnered with Dr. Michael Hausser's research group at the University College London to design a custom Bergamo microscope system to suit their needs. If you have additional questions about our Bergamo imaging systems, please click on the Contact Me button to the right to send us an email.

Options at a Glance

Five-Axis Movement of Rotating Bergamo Microscopes

Highlights

 

Rapid Volumetric Imaging Using Bessel Beams
In partnership with the Howard Hughes Medical Institute and Prof. Na Ji (University of California at Berkeley), Thorlabs offers a Bessel beam module for our Bergamo® multiphoton laser scanning microscope. In vivo volume imaging of neuronal activity requires both submicron spatial resolution and millisecond temporal resolution. While conventional methods create 3D images by serially scanning a diffraction-limited Gaussian beam, Bessel-beam-based multiphoton imaging relies on an axially elongated focus to capture volumetric images. The excitation beam’s extended depth of field creates a 2D projection of a 3D volume, effectively converting the 2D frame rate into a 3D volumetric rate. 

As demonstrated in Ji’s pioneering work, this rapid Bessel beam-based imaging technique has synaptic resolution, capturing Ca2+ dynamics and tuning properties of dendritic spines in mouse and ferret visual cortices. The power of this Bessel-beam-based multiphoton imaging technique is illustrated in the images below, which compare a 300 x 300 μm scan of a Thy1-GFP-M mouse brain slice imaged with Bessel (left) and Gaussian (right) scanning. 45 optical slices taken with a Gaussian focus are vertically stacked to generate a volume image, while the same structural features are visible in a single Bessel scan taken with a 45 μm-long focus. This indicates a substantial gain in volume-imaging speed, making this technique suitable for investigating sparsely labeled samples in-vivo.

If you are interested in upgrading your Bergamo microscope to include the Bessel beam imaging modality, please fill out our multiphoton microscope contact form or call (703) 651-1700. For a list of all the upgrades and add-ons available, please see the Retrofits tab.

Source: Lu R, Sun W, Liang Y, Kerlin A, Bierfeld J, Seelig JD, Wilson DE, Scholl B, Mohar B, Tanimoto M, Koyama M, Fitzpatrick D, Orger MB, and Ji N. "Video-rate volumetric functional imaging of the brain at synaptic resolution." Nature Neuroscience. 2017 Feb 27; 20: 620-628.

A single Bessel scan (left) captures the same structural information obtained from a Gaussian volume scan created by stacking 45 optical sections (right), reducing the total scan time by a factor of 45. The images show a brain slice scanned over a 300 μm x 300 μm area. Scan depth for the Gaussian stack is indicated by the scale bar. Sample Courtesy of Qinrong Zhang, PhD and Matthew Jacobs; the Ji Lab, Department of Physics, University of California, Berkeley.

 

3P Image
Click to Enlarge

A Thy1-YFP male mouse, 21 weeks old, imaged at 1300 nm, 326 kHz repetition rate, pulse width ~60 fs. At the top of the cortex (0 µm, 1.1 mW laser power), the window was centered at 2.5 mm lateral and 2 mm posterior from the Bregma point over somatosensory cortex. Courtesy of the Chris Xu Group, Cornell University.

Three-Photon Imaging
For our Bergamo multiphoton microscope, we have developed scan path optics for the 800 - 1800 nm range to open the door to three-photon techniques. Three-photon excitation is ideal for deep tissue imaging and requires a high-pulse-energy excitation source, typically around 1300 nm or 1700 nm. Compared to two-photon imaging, three-photon imaging offers less tissue scattering and reduced out-of-focus background, which results in an improved signal-to-background ratio.

Configurations capable of three-photon imaging, such as the one shown below, can include a dichroic mirror to support simultaneous two-photon and three-photon imaging, as well as electronics to support low-repetition-rate lasers with high bandwidth sampling. ThorImage®LS software has been enhanced with important features for three-photon detection. For instance, users can synchronize the three-photon signal detection to the excitation pulses and control the phase delay for peak signal-to-noise ratio. For more details, please see the ThorImageLS tab.

If you are interested in upgrading your Bergamo microscope to include the three-photon imaging modality, please fill out our multiphoton microscope contact form or call (703) 651-1700. For a list of all the upgrades and add-ons available, please see the Retrofits tab.

Source: Wang T and Xu C. "Three-photon neuronal imaging in deep mouse brain." Optica. 2020; 7 (8): 947-960.

Configuration for 2P and 3P
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Two- and Three-Photon Imaging Configuration. For more details, please see the Configurations tab.

 

Spatial Light Modulator for Simultaneous Multi-Site Activation
Thorlabs’ Spatial Light Modulator (SLM) uses holography patterns to enable photoactivation of multiple locations in a specimen simultaneously. Designed for two-photon excitation with femtosecond pulses, the SLM manipulates the phase across the stimulation laser beam profile to generate hundreds of user-determined focal points.

The diagrams below illustrate the benefits of using the SLM with two-photon activation over two-photon activation alone and single-photon activation. With single-photon activation, unintended nearby cells as well as the target cell become activated because this technique lacks the ability to target a single cell. This problem can be solved with two-photon activation, which allows single-cell resolution targeting; however, only one cell can be targeted at a time. Two-photon activation with SLM overcomes these limitations by generating a number of focal points and allowing multiple target cells to be activated simultaneously. Each beam can be shaped to improve the efficacy of photoactivation, a crucial feature for activating neural populations at varying depths within a single FOV. The SLM phase mask pattern can be rapidly switched, enabling multiple individual focal points to be targeted independently in any sequence. The calibration process, hologram generation, and external hardware synchronization are entirely managed through the ThorImage®LS software, enabling seamless control. For more details, please see the ThorImageLS tab.

If you are interested in upgrading your Bergamo microscope to include the SLM imaging modality, please fill out our multiphoton microscope contact form or call (703) 651-1700. For a list of all the upgrades and add-ons available, please see the Retrofits tab.

Two-Photon Activation + Spatial Light Modulator

  • Generate Multiple Focal Points
  • Simultaneously Activate Multiple Target Cells
Gaussian Scan
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Two-Photon Activation

  • Single-Cell Resolution Targeting
  • Activate Only Target Cell
Gaussian Scan
Click to Enlarge

Single-Photon Activation

  • Lack of Depth Control for 3D Neural Activation
  • Activate Target Cell and Unintended Nearby Cells
Gaussian Scan
Click to Enlarge

Two-photon activation with SLM (right) allows simultaneous excitation of multiple target cells, which is not possible with single-photon (left) or two-photon (middle) activation.

Features

Many of these features can also be added to existing Thorlabs microscope systems. For more information, please see the Retrofits tab.

Laser Scanning, Widefield Imaging, and Transmitted Light Imaging
Scan Paths Resonant-Galvo-Galvo
  • 8 kHz Scanner: Image at 2 fps (4096 x 4096 Pixels), 30 fps (512 x 512 Pixels), or 400 fps (512 x 32 Pixels)
  • 12 kHz Scanner: Image at 45 fps (512 x 512 Pixels) or 600 fps (512 x 32 Pixels)
  • Design Registered Under US Patent 10,722,977
Galvo-Resonant
  • 8 kHz Scanner: Image at 2 fps (4096 x 4096 Pixels), 30 fps (512 x 512 Pixels), or
    400 fps (512 x 32 Pixels)
  • 12 kHz Scanner: Image at 45 fps (512 x 512 Pixels) or 600 fps (512 x 32 Pixels)
Galvo-Galvo
  • User-Defined Scan Geometries: Squares, Rectangles, Circles, Ellipses, Lines, and Polylines
  • Capture Weak Signals with Long Dwell Time Integration
  • Consistent Dwell Times Across Field of View
  • 48 fps at 512 x 32 Pixels and 70 fps at 32 x 32 Pixels
Spatial Light Modulator
  • Simultaneous Multi-Site Photostimulation using Holographic Beam Control
  • Entirely Controlled Through ThorImage®LS Software
  • Precise Z-Axis Control of Photoexcited Locations
Widefield Viewing
  • Compatible with Thorlabs or Third-Party C-Mount-Threaded Scientific Cameras
  • Locate Areas of Interest without Unnecessary Laser Excitation
Epi-Illumination
  • Single-Filter-Cube or Six-Filter-Turret Epi-Illuminator Modules
  • Visualize Samples with Fluorescence or Reflected Light
  • Variety of Available Sources for Brightfield Illumination
    • Thorlabs' Mounted and Solis® LEDs
    • Optional Port for Industry-Standard Liquid Light Guides
Dodt Gradient Contrast and DIC Modules
  • User-Removable Modules Convert Microscope Between In Vivo and Slice Imaging
  • Widefield Imaging or Laser Scanning
  • Dodt Gradient Contrast: View Features in Tissue Slices
  • DIC: View Features in Thinner, Transparent Samples
Optical Performance
PMT Configuration
  • Choice of Signal Collection Angles to Accommodate Different Sample Depths along Epi Path
    • 8° (For Two PMTs)a
    • 10° (For up to Four PMTs)a
    • 14° (For Two PMTs)a
  • Option for Highly Sensitive Forward Fluorescence Detection Channels along Transmitted Path
    • 13° (For up to Two PMTs)a
  • GaAsP and Multialkali PMTs Available
  • All PMTs Available with or without Mechanical Shutters for Photoactivation Experiments
  • Angles Quoted for an Objective with a Ø20 mm Entrance Pupil
Minimal Distance Between Objective
and First Collecting Lens
  • Large Collection Angle for Multiphoton Fluorescence Emission
  • Increased Collection Efficiency
Periscopes
  • Maintain Laser Alignment and Optical Performance over Microscope's Entire Travel Range
Scan Paths Designed In-House
  • Super Broadband Correction Range of 450 - 1100 nm, 680 - 1300 nm, or 800 - 1800 nm
  • Optimized for Photostimulation, Two-Photon Imaging, or Three-Photon Imaging
  • Specifically Designed to Match the Low-Magnification, High-NA Objectives Popularly Used in Multiphoton Microscopy
  • Take Full Advantage of the Latest Widely Tunable Ti:Sapphire and OPO Systems
  • Fill up to a Ø20 mm Back Aperture Objective
  • Up to F.N. 20
Day-to-Day Usage
Several Software Packages
  • ThorImage®LS: Our Internally Developed, Open-Source Solution
  • Full SDK for LabVIEW and C++ Available Upon Request
  • Compatible with ScanImage
Touchscreen Controller
  • Touchscreen Shows Current Position of All Axes
  • Tap to Save and Retrieve Two Positions in Space
Easy-to-Reach Emission Filters and Dichroic Holders
  • Filters are Accessed from the Front of the Microscope and Take Less than 5 Minutes to Exchange
Input and Output Triggers
  • Use Electrical Signals to Synchronize All Your Equipment
  • Input Triggers Can Start a Single Series or an Indefinite Series
  • Output Triggers Can be Sent at the Beginning of a Frame or Line
  • High-Bandwidth Signal Integration with Electrophysiology
Large User-Adjustable Volume Underneath Objective
  • Accommodates Large Preps and Setups
  • Approach Angles Around Objective are Not Restricted
  • Rotating Bodies have 5" (12.7 cm) of Elevator Travel for Easily Adapting the Microscope for Differently Sized Experimental Setups
≥90° Rotation of the Focal Plane
(Rotating Bodies Only)
  • Image Different Sections of the Brain without Having to Move Your Specimen or Refocus the Objective
  • Rotation Angles:
    • 0° to 90° or -45° to +45° Rotation Around the Sample
Beam Conditioning Modules
Fast Laser Power Modulators
  • 0 to 20 kHz Modulation Frequency
  • Contrast Ratios of 250:1 or 2500:1 Available
  • Compatible with Galvo-Galvo Scanner Only
  • See Full Web Presentation Here
Pockels Cells
  • Edge and Fly-Back Blanking to Minimize Sample Photobleaching
  • Fast (1 MHz) and Slow (250 kHz) Masking for ROIs
  • Customize Laser Power at Each Slice Using Software Control
Variable Attenuator
  • Manual and Computer Control of Laser Power in Systems Without a Pockels Cell
  • Improves Pockels Cell Performance
  • One-Click Shutter
Beam Stabilizer
  • Maintain Stable Beam Pointing During Laser Excitation, Laser Wavelength Switching, and Temporal Drift
Volume Imaging Using Bessel Beams
  • 3D Video-Rate for Volumetric Functional Imaging
  • Enhanced Temporal Resolution Adequate for Studying Internal Systems at Cellular Lateral Resolution In Vivo
  • Press Release
Sample Holders
Rigid Stands for Slides, Recording Chambers, or Platforms
  • Minimal Footprint Conserves Space Around Objective and the Microscope
  • Slim Profile Leaves Room for Dodt or DIC Imaging Modules
  • Excellent Long-Term Stability
  • Easily Rotate Samples Into and Out of the Beam Path
XY Platforms Ideal for Micromanipulators
  • Large Working Space that Surrounds the Objective on Three Sides
  • Ideal for Setups Where the Sample and Apparatus Need to Move in Unison, Such as Patch Clamping
  • 2" Travel in X and Y; 0.5 µm Encoder Resolution
Thorlabs Support
Fully Designed and Manufactured In-House
  • Engineers Work Under One Roof to Lower Your Costs and Create Seamless Solutions
  • Expertise in Every System Component
Modular System Construction
  • As Your Experimental Needs Evolve, Upgrade Your Microscope Without Sacrificing Existing Capabilities
Professional Installation
  • Thorlabs Technician Visits Your Lab to Assemble, Test, and Demonstrate Use of Your Microscope
Quick Support
  • Thorlabs Technicians and Application Specialists Available for Videoconferencing
  • Communicate with Our Support Staff Faster than an Engineer Could Travel to Your Location
  • Thorlabs Will Ship You a Camera with a Microphone to Facilitate the Conversation
  • With Permission, Thorlabs Will Remote Desktop in to Address Software Issues

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Bergamo® II Modules

Thorlabs Bergamo® II microscopes are modular systems that can be customized in the design process to meet the exact needs of the experiment. The modules listed below are displayed in a variety of pre-built examples found on our Configurations tab to help provide a starting point for your design. 

 

BergamoII Rotating Body

Galvo-Resonant Scanners, Galvo-Galvo Scanners, and Spatial Light Modulators

Bergamo® II microscopes can be configured with one or two co-registered scan paths to propagate, condition, and direct an input laser beam. Each path can utilize a resonant-galvo-galvo scanner, galvo-resonant scanner, galvo-galvo scanner, and/or a spatial light modulator (SLM). These choices allow the user to optimize each experiment as needed for high frame rates, high sensitivity, and/or targeted exposure of the regions of interest.

Resonant-Galvo-Galvo Scanners for Multimodal Scanning
Thorlabs offers 8 kHz and 12 kHz resonant-galvo-galvo (RGG) scanners. The design of our RGG scanners is registered under US Patent 10,722,977. This multimodal scanner provides features of both the galvo-resonant and galvo-galvo scanners in a single scan head. Our 8 kHz scanners utilize the entire field of view and offer a maximum frame rate of 400 fps, while our 12 kHz scanners provide an increased frame rate of 600 fps.

Galvo-Resonant Scanners for High-Speed Imaging
Thorlabs offers 8 kHz and 12 kHz galvo-resonant scanners. Our 8 kHz scanners utilize the entire field of view and offer a maximum frame rate of 400 fps, while our 12 kHz scanners provide an increased frame rate of 600 fps.

Galvo-Galvo Scanners for User-Defined ROI Shapes
Galvo-galvo scanners support user-drawn scan geometries (lines, polylines, squares, and rectangles) and also support custom photoactivation patterns (circles, ellipses, polygons, and points). They offer consistent pixel dwell times for better signal integration and image uniformity.

Spatial Light Modulator for Simultaneous Targeting
Unlike scanners, which physically move from point to point, spatial light modulators (SLMs) use holography to diffract the beam and shape it in a user-defined pattern. This includes general beam shaping as well as the creation of multiple focal points at the FOV; the latter allows multiple sites in a sample to be photoexcited simultaneously.

Scan Paths of Example Configurations
B243
Multi-Target Photoactivation (Rotating) Path 1: Galvo-Resonant
Path 2: SLM
B242: Two- and Three-Photon Imaging (Rotating) Path 1: Galvo-Resonant
Path 2: Galvo-Galvo
B251: Random Access Scanning (Rotating) Resonant-Galvo-Galvo
B241:
In Vivo Two-Photon Imaging (Rotating) Galvo-Resonant
B252:
Dual-Path Random Access Scanning (XYZ) Path 1: Resonant-Galvo-Galvo
Path 2: Galvo-Galvo
B262:
Dual-Path Confocal Imaging (XYZ) Path 1: Galvo-Resonant
Path 2: Galvo-Galvo
B231:
Simple Imaging (XYZ) Galvo-Resonant
B211:
Video and High-Speed Imaging (Z-Only) Galvo-Galvo
B201:
Simple Imaging (Z-Only) Galvo-Galvo

 

Figure 3. Wavelength Switching Using Tiberius Ti:Sapphire Laser Shown at 1/16th Actual Speed
Fast Switching
Click to Enlarge
Figure 2. Fast Switching between the optimal excitation wavelengths of 750 nm and 835 nm provides the high contrast seen in this composite image. The two-channel set was collected at an imaging rate of 7 fps.
Fast Switching
Click to Enlarge
Figure 1.  The above image was acquired using single-wavelength excitation at 788 nm, while the optimum excitation wavelengths for the two tags are 750 nm and 850 nm.

Fast Switching Using a
Tunable Femtosecond Laser

With an industry-leading tuning speed of up to 4000 nm/s and a wide 720 to 1060 nm tuning range, the Tiberius® Ti:Sapphire Femtosecond Laser is ideal for fast sequential imaging in multiphoton microscopy applications. 

A 25 µm thick sagittal section of an adult rat brain is shown in the images and video to the right. The red channel corresponds to fluorescence from chick anti-neurofilament that is optimally excited at 835 nm, while the green channel corresponds to fluorescence from mouse anti-GFAP that is optimally excited at 750 nm.

Figure 1 shows fluorescence from single-wavelength excitation at 788 nm, which sub-optimally excites the two tags simultaneously. Figure 2 is a composite image of the fluorescence produced by a two-color excitation image sequence acquired at 7 fps where the excitation wavelength was rapidly tuned between 750 and 835 nm. The video in Figure 3 shows the fast-switching used to create the composite image in Figure 2 at 1/16th of the actual speed. When compared to single-wavelength excitation at 788 nm with the same intensity, fast switching offers much higher image contrast as it provides optimal excitation of both fluorophores.

This immunofluorescence sample was prepared by Lynne Holtzclaw of the NICHD Microscopy and Imaging Core Facility, a part of the National Institutes of Health (NIH) in Bethesda, MD.

 


Click to Enlarge
Figure 2. Close-Up of a Gaussian Beam

Click to Enlarge
Figure 1. Close-Up of a Bessel Beam

Volumetric Imaging Technique Using Bessel Beams

Thorlabs is excited to offer a new ultra-fast imaging technique that uses a Bessel beam to provide video-rate volumetric functional imaging of neuronal pathways and interactions in vivo. These unique beams are non-diffractive and self-healing, which allows them to maintain a tight focus and even reform as they pass through tissue. This technique is offered for Thorlabs' Bergamo II multiphoton microscopes and Thorlabs' Multiphoton Mesoscope

The images to the right depict a Bessel beam and a Gaussian beam, respectively. As you can see in the images, the Gaussian beam has a singular point of focus that progressively becomes weaker as it diverges from the central point, whereas the Bessel beam has a beam annulus that maintains its focus.

To read the press release about this new technique, click here.

 

BergamoII Rotating Body
Click for Details

Rotating Bergamo II systems are outfitted with multi-joint articulating periscopes. This periscope's design offers the enhanced flexibility needed to allow the entire scanning system to be tilted with respect to the sample.
BergamoII Non-Rotating Body
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Upright Bergamo II systems are equipped with periscopes that permit the microscope's full travel range in X, Y, and Z to be used without compromising the optical performance.

Periscopes

Most lasers used in multiphoton microscopy are delivered by a free-space beam. The Bergamo II's ability to translate the objective around the focal plane in up to four axes (X, Y, Z, and θ) also requires the beam path to translate along the same axes while maintaining alignment. Bergamo II systems overcome this engineering challenge using multi-jointed periscopes.

Configurations with Articulating Periscopes
B243: Multi-Target Photoactivation (Rotating)
B242:
Two- and Three-Photon Imaging (Rotating)
B251: Random Access Scanning (Rotating)
B241: In Vivo Two-Photon Imaging (Rotating)
Configurations with Fixed Periscopes
B252: Dual-Path Random Access Scanning (XYZ)
B262:
Dual-Path Confocal Imaging (XYZ)
B231: Simple Imaging (XYZ)
B211:
Video and High-Speed Imaging (Z-Only)
B201:
Simple Imaging (Z-Only)

 

BergamoII Filter Exchange
Click to Enlarge

Emission filters and dichroic cubes are held behind magnetically sealed doors on the front of the PMT detection module.
Collection Optics of Example Configurationsa
B243:
Multi-Target Photoactivation (Rotating) Epi: 10°
B242: Two- and Three-Photon Imaging (Rotating) Epi: 14°
B251: Random Access Scanning (Rotating) Epi: 14°
B241: In Vivo Two-Photon Imaging (Rotating) Epi: 14°
B252: Dual-Path Random Access Scanning (XYZ) Epi: 14°
Transmitted: 13°
B262: Dual-Path Confocal Imaging (XYZ) Epi: 14°
B231: Simple Imaging (XYZ) Epi: 8°
Transmitted: 13°
B211: Video and High-Speed Imaging (Z-Only) Epi: 
B201: Simple Imaging (Z-Only) Epi: 

Super Broadband Scan Optics

Bergamo II microscopes feature proprietary scan optics that are optimized and corrected for excitation wavelengths within the 450 - 1100 nm, 680 - 1300 nm, or 800 - 1800 nm wavelength range, ideal for photostimulation, two-photon imaging, and three-photon imaging, respectively. These broad ranges, extending from the visible well into the near infrared, were chosen to support the latest widely tunable Ti:Sapphire lasers and OPO systems, as well as dual-output lasers such as the Chameleon Discovery.

Our optics take full advantage of the optical designs used in the low-magnification, high-numerical-aperture objectives by filling the back aperture of the objective up to Ø20 mm. This creates an exceptional scan area that lets you find a region of interest more quickly or simply image more cells at once.

Large-Angle Signal Collection Optics

Deriving the most signal from limited photons is the fundamental goal of any detection system. By positioning the PMTs immediately after the objective (a "non-descanned" geometry), light that is scattered by the sample, which therefore appears to originate outside the objective's field of view, still strikes the PMTs and adds to the collected signal. This is a benefit unique to multiphoton microscopy. Collecting beyond the objective's design field of view greatly enhances overall detection efficiency when imaging deep in tissue.

In the epi direction, we offer signal collection anglesa of 8°, 10°, or 14°, while in the transmitted direction, we offer a signal collection anglea of 13°. Our collection modules can optionally be outfitted with mechanical shutters for photoactivation experiments.

Easy-to-Reach Emission Filters and Dichroic Holders

Bergamo II systems are fully compatible with industry-standard fluorescence filter sets that include Ø25 mm fluorescence filters and 25 mm x 36 mm dichroic mirrors. Unlike competing designs, Thorlabs' detector modules have magnetic holders that make it simple and quick to exchange filters for different measurements.

We also offer detection modules for large-area Ø32 mm fluorescence filters and 32 mm x 44 mm dichroics, which support greater collection angles for increased signal.

Detectors in Epi and Transmitted Directions

We employ high-sensitivity GaAsP PMTs in our multiphoton systems, which can offer high quantum efficiency, aiding in imaging weakly fluorescent or highly photosensitive samples. Our PMTs can either be thermoelectrically cooled for improved sensitivity toward weak signals or non-cooled for a smaller package size and greater numerical aperture. Multialkali PMTs are also available.

All Bergamo® II microscopes can be equipped with either two or four detection channels in the epi direction, and/or two detection channels in the forward direction. The user can configure the forward-direction channels to detect the same fluorescent tags as the epi-direction PMTs, raising the microscope's sensitivity toward thin, weakly fluorescent specimens.

A maximum of four channels can be controlled by the software at a given time.

  • Angles Quoted for an Objective with a Ø20 mm Entrance Pupil

 

Multi-Axis Controller with Touchscreen

This controller is specifically designed for rotating Bergamo II microscope bodies. It uses knobs to control up to five motorized axes. On rotating systems, a rocker switch changes between fine objective focusing and translation of the elevator base. Each axis can be disabled on an individual basis in order to maintain a location along the desired direction.

The integrated touchscreen lets two spatial locations be saved and retrieved locally. Up to eight spatial locations can be saved on the computer running ThorImage®LS. The touchscreen also reads out the position of every motor.

 

Objectives

Bergamo II microscopes accept infinity-corrected objectives with M34 x 1.0, M32 x 0.75, M25 x 0.75, or RMS threads. Together, these options encompass the majority of low-magnification, high-NA objectives used in multiphoton microscopy. With a large field number of 20, our scan optics completely utilize the optical designs of these specialized objectives, offering enhanced light-gathering ability compared to competing microscopes using the same objectives.

 

Rigid Stand Sample Holders

Thorlabs' Rigid Stands are rotatable, lockable, low-profile platforms for mounting slides, recording chambers, our Z-axis piezo stages, and custom experimental apparatuses. Each fixture is supported by a solid Ø1.5" stainless steel post for passive vibrational damping, which is in turn held to the workstation by the red post holder.

A locking collar maintains the height of the platform, allowing it to easily rotate into and out of the optical path, and a quick-release mechanism holds the post in place once the desired position is achieved.

 

Quantulux and 1.4 Megapixel Scientific Cameras
Click to Enlarge

A Zelux sCMOS Camera and a Quantulux sCMOS Camera

Scientific Cameras

Our low-noise sCMOS and CMOS cameras were designed for full compatibility with Thorlabs’ multiphoton microscopy systems. Useful for widefield and fluorescence microscopy, they are capable of visualizing in vitro and in vivo samples using reflected light and fluorescence emission. They work in conjunction with the epi-fluorescence module to help locate fiducial markers, and they also enable imaging modalities that do not require laser exposure.

Thorlabs' cameras are driven by our internally developed ThorCam software package. The sCMOS camera is available with a 2.1 MP sensor, and our CMOS cameras are available with either a 1.3 MP, 2.3 MP, 5 MP, 8.9 MP, or 12.3 MP sensor. Generally speaking, cameras with lower resolution offer higher maximum frame rates. These cameras also feature a separate auxiliary port that permits the image acquisition to be driven by an external electrical trigger signal.

Bergamo® II microscopes are also directly compatible with any camera using industry-standard C-mount or CS-mount threads.

 

Bergamo II with Dodt Contrast
Click to Enlarge
Trans-Illumination Module, Motorized Condenser Stage, and
Rigid Stand Sample Holder Underneath the Objective

User-Installable Dodt Contrast and DIC Imaging Modules

The modular construction of the Bergamo® II makes it exceptionally easy for the user to convert the microscope between in vitro and in vivo applications. Our user-installable trans-illumination modules for Dodt contrast, laser-scanned Dodt contrast, and differential interference contrast (DIC) take less than 5 minutes to attach or remove from the microscope body. These modules are available for both rotating and upright bodies.

Each option is paired with our basic 3-axis controller, which optimizes the illumination conditions by translating our motorized condenser stage over a 1" range. This versatile design is compatible with air and high-NA oil immersion condensers designed by Nikon.

To complement these modules, we manufacture slim-profile rigid stand sample holders that are ideal for positioning slides between the transmitted light module and the objective.

Configurations with Trans-Illumination Module
B252:
Dual-Path Random Access Scanning (XYZ)
B231: Simple Imaging (XYZ)

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Thorlabs' Bergamo® II Series Multiphoton Microscopy Platform is a powerful tool adapted to meet experimental needs across a wide range of research fields. Click on the images below to explore how a Bergamo can and has been utilized for each application.


Structural Neurobiology


Neurological Disorders


Neural Development and Plasticity


Neurogenetics


Functional and Molecular Imaging


Synapses and Circuits

Ion Channels, Transporters, and Neurotransmitter Reporters


Cell Biology of Neurons, Muscles, and Glia


Drug Discovery

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Top: Astrocytes are labelled with SR101 (red). Arrows point to astrocytes that had Ca2+ elevations during tDCS. The numbers correspond to the cells and neurogliopil regions plotted in the graphs below. Bottom: Fluorescent intensity (ΔF/F) traces of astrocytes (orange), neurons (green) and neurogliopil (brown). Figure Courtesy of Monai H. et al. (See Below)

Application Summary

The study of cell biology, muscles, and glia often involves imaging in vitro or in vivo samples tagged with multiple fluorophores. Through multiphoton imaging with these fluorescent markers, researchers are able to observe gene expression related to neural function in different areas of the brain. We recommend two of our configurations for this application, see the table below. With two to four channel detection modules and fast sequential imaging using our Tiberius® Tunable fs laser, both of these configurations offer the flexibility necessary for experiments in this field. In addition, each configuration has a small footprint and a large throat depth to provide ample room for numerous sample mounting options, including in vivo imaging of mammalian brains via transcranial windows.

Body Upright XYZ Upright Z-Axis
Function Simple XYZ Imaging Video and High-Speed Imaging
Configuration B231 B211

Publications

Rose T, Jaepel J, Hübener M, and Bonhoeffer T. "Cell-specific restoration of stimulus preference after monocular deprivation in the visual cortex." Science. 2016 Jun 10; 352 (6291): 1319-1322.

Monai H, Ohkura M, Tanaka M, Oe Y, Konno A, Hirai H, Mikoshiba K, Itohara S, Nakai J, Iwai Y, and Hirase H. "Calcium imaging reveals glial involvement in transcranial direct current stimulation-induced plasticity in mouse brain." Nature Communications. 2016 March 22; 7: 11100.

Lu W, Xie Z, Tang Y, Bai L, Yao Y, Fu C, and Ma G. "Photoluminescent mesoporous silicon nanoparticles with siccr2 improve the effects of mesenchymal stromal cell transplantation after acute myocardial infarction." Theranostics. 2015 Jun 25; 5 (10): 1068-1082.

Qin X, Qiu C, and Zhao L. "Maslinic acid protects vascular smooth muscle cells from oxidative stress through Akt/Nrf2/HO-1 pathway." Molecular and Cellular Biochemistry. 2014 Feb 20; 390 (1-2): 61-67.

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Artistic Rendering of DNA

Application Summary

Drug discovery research is rapidly expanding and often requires a wide variety of experimental setups and imaging techniques. Two-photon imaging is frequently used to measure the characteristics of drug applications, including depth of drug penetration and the area of its spread throughout the cortex. We offer two configurations that are suitable for this application, see the table below. These configurations are are well-balanced for both in vitro and fixed stage in vivo microscopy research. The modularity of the Bergamo systems' removable trans-illumination module provides versatility in regard to experimental techniques, imaging modalities, and sample subjects.

Body Upright XYZ
Function Simple XYZ Imaging Bessel Beam
Configuration B231 B252

Publications

Strobl MJ, Freeman D, Patel J, Poulsen R, Wendler CC, Rivkees SA, and Coleman E. " Opposing effects of maternal hypo- and hyperthyroidism on the stability of thalamocortical synapses in the visual cortex of adult offspring." Cerebral Cortex. 2017 May 1; 27 (5): 3015-3027.

Scattolini V, Luni C, Zambon A, Galvanin S, Gagliano O, Ciubotaru CD, Avogaro A, Mammano F, Elvassore N, and Fadini GP. "Simvastatin rapidly and reversibly inhibits insulin secretion in intact single-islet cultures." Diabetes Therapy. 2016 Nov 9; 7 (4): 679-693.

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Imaging Visually Evoked Synaptic Calcium Transient In Vivo, Using Dendrite Labels with GCAMP6, 200 µm Deep in Layer 2/3 Visual Cortex, Courtesy of David Fitzpatrick, Max Plank Institute for Neurobiology, Jupiter, FL, USA

Application Summary

In vivo functional imaging of organisms and the individual neurons linked to specific organism behaviors requires high-speed and high-sensitivity imaging. We offer six configurations for this application (see the table below). These configurations are equipped with an 8 or 12 kHz galvo-resonant imaging scanner and our 10° or 14° wide-angle collection optics to enable fast, high-resolution imaging. Each microscope system features a large throat depth, 5” of vertical travel, and smooth movement along the Z-axis to create a large working space ideal for in vivo volume imaging deep into highly scattering samples of neural tissue. For an improved range of movement around a sample, we recommend a configuration with a rotating body.

Body Rotating Upright XYZ Upright
Z-Axis
Function Multi-Target Photoactivation Dual-Modality
and
In Vivo
Imaging
In Vivo Two-Photon Imaging Dual-Path with Confocal Imaging Simple XYZ Imaging Video and High-Speed Imaging
Configuration B243 B251 B241 B262 B231 B211

Publications

Aghayee S, Bowen Z, Winkowski DE, Harrington M, Kanold P, and Losert W. "Particle tracking facilitates real time motion compensation in 2D or 3D two-photon imaging of neuronal activity." Frontiers in Neural Circuits. 2017 Aug 15; 11: 56.

Schnell B, Ros IG, and Dickinson MH. "A descending neuron correlated with the rapid steering maneuvers of flying Drosophila." Current Biology. 2017 Apr 6; 27 (8): 1200-1205.

Roth MM, Dahmen JC, Muir DR, Imhof F, Martini FJ, and Hofer SB. "Thalamic nuclei convey diverse contextual information to layer 1 of visual cortex." Nature Neuroscience. 2015 Dec 21; 19: 299-307.

Barnstedt O, Keating P, Weissenberger Y, King AJ, and Dahmen JC. "Functional microarchitecture of the mouse dorsal inferior colliculus revealed through in vivo two-photon calcium imaging." The Journal of Neuroscience. 2015 Aug 5; 35 (31): 10927-10939.

Boyd AM, Kato HK, Komiyama T, and Isaacson JS. "Broadcasting of cortical activity to the olfactory bulb." Cell Reports. 2015 Feb 24; 10 (7): 1032-1039.

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Cochlear Organotypic Culture Loaded with Fluo4-AM and DM-Nitrophen AM, Calcium Release Triggered Using Galvo-Galvo Uncaging Pathway in Outer Hair Cell Indicated by Red Box After ~13 Seconds, Courtesy of Federico Ceriani and Walter Marcotti, University of Sheffield

Application Summary

In this application, scientists are interested in neural connections and intercellular movement. With multiphoton imaging, they are able to trace the direction and speed of ions moving through channel membrane proteins or neurotransmitters moving from one neuron to another. This research requires a microscopy system that has high-resolution and high-speed imaging of multiple fields of view (FOVs). We recommend six of our multiphoton configurations for this application (see the table below). This area of research often requires both in vivo and in vitro imaging within the same study, so within each configuration, our transmitted light modules can be installed or removed by the user in just a few minutes, making it exceptionally easy to switch between the two imaging modalities. These configurations are capable of high-frame-rate imaging and targeted laser activation for photostimulation, making them ideal for correlating neural responses in multiple regions of the brain.

Body Rotating Upright XYZ Upright
Z-Axis
Function Multi-Target Photoactivation Dual-Modality and In Vivo Imaging In Vivo Two-Photon Imaging Bessel Beam Dual-Path with Confocal Imaging Video and High-Speed Imaging
Configuration B243 B251 B241 B252 B262 B211

Publications

De Toni L, Garolla A, Menegazzo M, Magagna S, Di Nisio A, Šabovic I, Rocca MS, Scattolini V, Filippi A, and Foresta C. "Heat sensing receptor TRPV1 is a mediator of thermotaxis in human spermatozoa." PLoS One. 2016 Dec 16; 11 (12): e0167622.

Mongeon R, Venkatachalam V, and Yellen G. "Cytosolic NADH-NAD+ redox visualized in brain slices by two-photon fluorescence biosensor imaging." Antioxidants & Redox Signaling. 2016 Oct 1; 25 (10): 553-563.

Lewin AE, Vicini S, Richardson J, Dretchen KL, Gillis RA, and Sahibzada N. "Optogenetic and pharmacological evidence that somatostatin-GABA neurons are important regulators of parasympathetic outflow to the stomach." The Journal of Physiology. 2016 Mar 9; 594 (10): 2661-2679.

Cossell L, Iacaruso MF, Muir DR, Houlton R, Sader EN, Ko H, Hofer SB, and Mrsic-Flogel TD. "Functional organization of excitatory synaptic strength in primary visual cortex." Nature. 2015 Feb 19; 518: 399-403.

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Laser-Scanned Two-Photon SHG+Dodt Gradient Contrast of Zebrafish Embryo

Application Summary

Neurogenetic studies require a wide variety of experimental setups for in vivo research. Multiphoton imaging is ideal for studying live organisms, especially embryos, as it reduces the occurrence of photobleaching and phototoxicity that is common with other light microscopy techniques. There are three multiphoton configurations we suggest for Neurogenetic applications (see the table below). The small footprint and large throat depth of each system provide ample room for sample mounts and experimental apparatus, such as the large setup used for Drosophilia studies by Chen et al. (click here for supplementary videos). Additionally, these systems provide video-rate, sequential two-photon imaging to study fast dynamic biological and chemical processes in vivo without damaging the sample.

Body Upright XYZ Upright Z-Axis
Function Simple XYZ Imaging Video and High-Speed Imaging Simple Z-Axis Imaging
Configuration B231 B211 B201

Publications

Chen CL, Hermans L, Viswanathan MC, Fortun D, Aymanns F, Unser M, Cammarato A, Dickinson MH, and Ramdya P. "Imaging neural activity in the ventral nerve cordof behaving adult Drosophila." Nature Communications. 2018 October 22; 9: 4390.

Dechen K, Richards CD, Lye JC, Hwang JEC, and Burke R. "Compartmentalized zinc deficiency and toxicities caused by ZnT and Zip gene over expression result in specific phenotypes in Drosophila." The International Journal of Biochemistry & Cell Biology. 2015 Jan 3; 60: 23-33.

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Two-Photon Image of Neurons Expressing Thy1-YFP in a Cleared Region of the Hippocampus, Courtesy of the 2017 Imaging Structure and Function in the Nervous System Course at Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

Application Summary

Scientists interested in this application focus on tracing neuronal pathways and researching dendritic spine plasticity. Imaging systems used in this type of research are capable of two-photon calcium imaging and/or confocal fluorescence imaging. High-resolution and high-sensitivity are crucial features for these systems. We offer three multiphoton configurations that are ideal for this application (see the table below). Each configuration may be used in photon-limited environments as they feature sensitive detection modules, in addition to our 10° or 14° full field-of-view collection optics with ultrasensitive GaAsP PMTs. Our Dual-Path with Confocal Imaging configuration allows for a range of experimental conditions to be observed using any combination of multiphoton, confocal, and epi-fluorescence imaging, along with photoactivation. Alternatively, the Video and High-Speed Imaging and Simple Z-Axis Imaging configurations provide a small footprint and large throat depth for in vivo two-photon imaging.

Body Upright XYZ Upright Z-Axis
Function Dual Path with Confocal Imaging Video and High-Speed Imaging Simple Z-Axis Imaging
Configuration B262 B211 B201

Publications

Gillet SN, Kato HK, Justen MA, Lai M, and Isaacson JS. "Fear learning regulates cortical sensory representations by suppressing habituation." Frontiers in Neural Circuits. 2018 Jan 10; 11: 112.

Micu I, Brideau C, Lu L, and Stys PK. "Effects of laser polarization on responses of the fluorescent Ca2+ indicator X-Rhod-1 in neurons and myelin." Neurophotonics. 2017 Jun 5; 4 (2): 025002.

Chen SX, Kim AN, Peters AJ, and Komiyama T. "Subtype-specific plasticity of inhibitory circuits in motor cortex during motor learning." Nature Neuroscience. 2015 Jun 22; 18: 1109-1115.

Peters AJ, Chen SX, and Komiyama T. "Emergence of reproducible spatiotemporal activity during motor learning." Nature. 2014 Jun 12; 510: 263-267.

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Stitched Confocal Fluorescence Image of Rat Retina Stained with DAPI, Alexa Fluor® 555 and Alexa Fluor® 633, Courtesy of Dr. Jennifer Kielczewski, National Eye Institute, National Institutes of Health, Bethesda, MD

Application Summary

Researching neurological disorders involves measuring neural function using two-photon calcium imaging. This research requires fast image acquisition and photostimulation. We recommend three of our multiphoton microscope configurations for this application (see the table below). Each configuration offers a large working space and rotating microscope body, making it ideal for in vivo animal studies. Our Multi-Target Photoactivation configuration features a spatial light modulator (SLM), which allows the activation of groups of neurons at varying depths within a single field of view. For increased penetration of samples with scattering tissues, the three-photon capability of our Two- and Three-Photon Imaging configuration is recommended. Our Dual-Modality and In Vivo Imaging configuration uses a resonant-galvo-galvo scanner to take high-resolution images. This scanner provides all the speed of a resonant-galvo scanner, while enabling user-defined photoactivation regions.

Body Rotating
Function Multi-Target Photoactivation Two- and Three-Photon Imaging Dual-Modality and
In Vivo Imaging
Configuration B243 B242 B251

Publications

Murphy SC, Palmer LM, Nyffeler T, Müri RM, and Larkum ME. "Transcranial magnetic stimulation (TMS) inhibits cortical dendrites." eLife. 2016 Mar 18; 5: e13598.

Lalchandani RR, van der Goes MS, Partridge JG, and Vicini S. "Dopamine D2 receptors regulate collateral inhibition between striatal medium spiny neurons." The Journal of Neuroscience. 2013 Aug 28; 33 (35): 14075-14086.

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1.2 mm In Vivo Deep Brain 3D Image Stack, Courtesy of Dr. Hajime Hirase and Katsuya Ozawa, RIKEN Brain Science Institute, Wako, Japan

Application Summary

Scientists investigate the structure of the brain to understand functions of neuronal proteins as well as the causes of neurological diseases. Due to the difficulty of imaging through brain tissue, this study often requires a multi-modality configuration allowing for a range of experimental conditions using any combination of multiphoton, confocal, and epi-fluorescence imaging. The three example multiphoton microscope configurations in the table below are designed to accommodate the needs of Structural Biology. Each configuration features fast-Z power ramping to accomplish high-resolution imaging deep within a sample. Our Two- and Three-Photon Imaging configuration uses both galvo-resonant and galvo-galvo scanners and infrared wavelength scanning optics to image second- and third-harmonic generation (SHG and THG). Alternatively, our Dual-Path Multiphoton Microscope with Confocal Imaging is outfitted with a confocal path that accommodates up to 4 laser lines and a 4-channel PMT detection module. The addition of a six-cube epi-illuminator module and sCMOS Quanatlux camera allows this system to perform epi-fluorescence imaging. Our Simple XYZ Imaging configuration is well-balanced for both in vitro and fixed stage in vivo microscopy research. With a removable transmitted illumination module, this versatile system can support a wide variety of experimental techniques, imaging modalities, and sample subjects.

Body Rotating Upright XYZ
Function Two- and Three-Photon Imaging Dual Path with Confocal Imaging Simple XYZ Imaging
Configuration B242 B262 B231

Publications

Lee KS, Huang X, and Fitzpatrick D. "Topology of ON and OFF inputs in visual cortex enables an invariant columnar architecture." Nature. 2016 May 5; 533: 90-94.

Lang X and Lyubovitsky JG. "Structural dependency of collagen fibers on ion types revealed by in situ second harmonic generation (SHG) imaging Ion channels and G coupled receptors." Analytical Methods. 2014 Nov 13; 7 (5): 1637-2230.

Ehmke T, Nitzsche TH, Knebl A, and Heisterkamp A. "Molecular orientation sensitive second harmonic microscopy by radially and azimuthally polarized light." Biomedical Optics Express. 2014 Jul 1; 5 (7): 2231-2246.

Lang X, Spousta M, Hwang YJ, and Lyubovitsky JG. "Noninvasive imaging of embryonic stem cell cultures by multiphoton microscopy reveals the significance of collagen hydrogel preparation parameters." Analytical Methods. 2011 Jan 20; 3 (3): 529-536.

Cai F, Yu J, Qian J, Wang Y, Chen Z, Huang J, Ye Z, and He S. "Use of tunable second-harmonic signal from KNbO3 nanoneedles to find optimal wavelength for deep-tissue imaging." Laser & Photonics Review. 2014 Jun 17; 8 (6): 865-874.

Poguzhelskaya E, Artamonov D, Bolshakova A, Vlasova O, and Bezprozvanny I. "Simplified method to perform CLARITY imaging." Molecular Neurodegeneration. 2014 May 26; 9 (19).

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Simultaneous Photostimulation of 100 Cells Co-Expressing GCaMP6f (Green) and C1V1 (Red), Courtesy of Lloyd Russell, Dr. Adam Packer, and Professor Michael Häusser, University College London, United Kingdom

Application Summary

By studying synapses and circuits, researchers are able to understand neuronal activity. Imaging synapses and circuits often requires simultaneous stimulation of populations of neurons. To achieve this, we offer three configurations of our multiphoton microscope capable of fast image acquisition of multiple regions within a single field of view (see table below). Our Multi-Target Photoactivation configuration features a spatial light modulator (SLM), which allows multiple sites in a sample to be photoexcited simultaneously. With the SLM, each beamlet can be shaped to improve the efficacy of photoactivation, a crucial feature for activating neural populations at varying depths within a single field of view (FOV). Our Dual-Modality and In Vivo Imaging configuration has a resonant-galvo-galvo scanner which provides features of both the galvo-resonant and galvo-galvo scanners in a single scan head. This scanner provides all the speed of a resonant-galvo scanner for high-resolution images, while enabling user-defined photoactivation regions. Lastly, our In Vivo Two-Photon Imaging configuration uses a galvo-resonant scanner for high-speed imaging.

Body Rotating
Function Multi-Target Photoactivation Dual-Modality and
In Vivo Imaging
In Vivo Two-Photon Imaging
Configuration B243 B251 B241

Publications

Scholl B, Wilson DE, and Fitzpatrick D. "Local order within global disorder: synaptic architecture of visual space." Neuron. 2017 Dec 6; 96 (5): 1127-1138.

Leinweber M, Ward DR, Sobczak JM, Attinger A, and Keller GB. "A sensorimotor circuit in mouse cortex for visual flow predictions." Neuron. 2017 Sep 13; 95 (6): 1420-1432.

Iacaruso, MF, Gasler IT, and Hofer SB. "Synaptic organization of visual space in primary visual cortex." Nature. 2017 Jul 27; 547: 449-452.

Dipoppa M, Ranson A, Krumin M, Pachitariu M, Carandini M, and Harris KD. "Vision and locomotion shape the interactions between neuron types in mouse visual cortex." Neuron. 2016 May 2; 98 (3): 602-615.

Example Configurations

Explore the details of example Bergamo II rotating, XYZ, and Z-axis system configurations by clicking on the expandable sections below. The modular nature of our multiphoton microscopy platform allows us to modify configurations to meet individual experimental needs or adjust the functionality of a microscope after installation; more information on modules featured in the systems below can be found on the Modules tab.

Configuration System Highlights
Rotating Body
Configuration B243:
Multi-Target Photoactivation
Key Features Suggested Applications
  • Spatial Light Modulator (SLM) in Secondary Beam Path, Providing Holographic, All-Optical Multi-Target Stimulation
  • Large Throat Depth for In Vivo Animal Studies
  • Separate Lasers for Multiphoton Imaging and Photoactivation
  • Galvo-Resonant Scanning for High-Speed Image Acquisition
  • Synapses and Circuits
  • Ion Channels, Transporters, and Neurotransmitter Reporters
  • Functional and Molecular Imaging
  • Neurological Disorders
Configuration B242:
Two- and Three-Photon Imaging
Key Features Suggested Applications
  • Dual-Channel Paths for Simultaneous 2P and 3P Imaging, or 3P Imaging with Photoactivation
  • Beam Conditioning with Pockels Cell and Laser Attenuator
  • Simultaneous 2P or 3P Imaging with Widefield
    Epi-Fluorescence
  • Large Working Space and Rotating Microscope Body for In Vivo Large Animal Studies
  • Structural Neurobiology
  • Neurological Disorders
Configuration B251:
Dual-Modality and
In Vivo Imaging
Key Features Suggested Applications
  • Resonant-Galvo-Galvo Scanner (US Patent 10,722,977) for High-Speed, High-Resolution Imaging
  • Integrate with Soundbox for Sound-Sensitive Experiments
  • Edge Blanking with Fast Pockels Cell
  • Simultaneous Multi-Channel Epi-Fluorescence
  • Tiberius fs Ti:Sapphire Tunable Laser
  • Synapses and Circuits
  • Ion Channels, Transporters, and Neurotransmitter Reporters
  • Functional and Molecular Imaging
  • Neurological Disorders
Configuration B241:
In Vivo Two-Photon Imaging
Key Features Suggested Applications
  • Large Working Space with Fully Rotatable Microscope
  • Integrate with Soundbox for Sound-Sensitive Experiments
  • Galvo-Resonant Scanner for High-Speed Imaging
  • Edge Blanking with Fast or Standard Pockels Cell
  • Simultaneous Multi-Channel Epi-Fluorescence
  • Tiberius® fs Ti:Sapphire Tunable Laser
  • Ion Channels, Transporters, and Neurotransmitter Reporters
  • Functional and Molecular Imaging
  • Synapses and Circuits
  • Auditory Functional Imaging
Upright XYZ Body
Configuration B252:
Bessel Beam
Key Features Suggested Applications
  • Light-Tight Enclosure for Laser Conditioning Modules and Components
  • Resonant-Galvo-Galvo Scanner (US Patent 10,722,977) for High-Speed, High-Resolution Imaging
  • PMT Options for Scanned or De-Scanned Multiphoton Detection
  • Breadboard System for Samples and Supplementary Equipment
  • Dual-Output Spectra Physics InSight Laser
  • High-Speed, High-Resolution Volumetric Imaging Using Bessel Beams (Optional)
  • Ion Channels, Transporters, and Neurotransmitter Reporters
  • Drug Discovery
  • Ex Vivo Neurobiological Studies
  • Electrophysiology and Patch-Clamp Recordings
  • In Vivo Neurobiological Studies using Bessel Beam Volumetric Imaging Technique (Optional)
Configuration B262:
Dual-Path with
Confocal Imaging
Key Features Suggested Applications
  • Dual-Paths for Multiphoton Imaging with
    Confocal Imaging or Photoactivation
  • Galvo-Resonant Scanner for High-Speed Imaging and Galvo-Galvo Scanner for Custom Geometric Scans
  • Four-Channel Confocal Fiber Laser and Tiberius® fs Ti:Sapphire Tunable Laser
  • Structural Neurobiology
  • Ion Channels, Transporters, and Neurotransmitter Reporters
  • Neural Development and Plasticity
  • Functional and Molecular Imaging
Configuration B231:
Simple XYZ Imaging
Key Features Suggested Applications
  • High-Speed Galvo-Resonant Scanning
  • High-Sensitivity GaAsP PMT
  • Free-Space Photodetector
  • Removable Transmitted Light Module with Laser Scanning Dodt Functionality
  • Epi-Fluorescence with Quantalux™ sCMOS Camera
  • Tiberius fs Ti:Sapphire Tunable Laser
  • Structural Neurobiology
  • Functional and Molecular Imaging
  • Drug Discovery
  • Fixed Stage Experiments
  • Neurogenetics
  • Cell Biology of Neurons, Muscles, and Glia
Upright Z-Axis Body
Configuration B211:
Video and High-Speed Imaging
Key Features Suggested Applications
  • High-Speed Galvo-Resonant Scanning
  • Motorized Laser Attenuator for Remote Power Adjustment
  • 2-Channel PMT Detection
  • Small Footprint with Large Throat Depth
  • Tiberius fs Ti:Sapphire Tunable Laser
  • Neural Development and Plasticity
  • Neurogenetics
  • Ion Channels, Transporters, and Neurotransmitter Reporters
  • Functional and Molecular Imaging
  • Cell Biology of Neurons, Muscles, and Glia
Configuration B201:
Simple Z-Axis Imaging
Key Features Suggested Applications
  • Galvo-Galvo Scanning
  • 2-Channel Detection with High-Sensitivity GaAsP PMTs
  • Small Footprint with Large Throat Depth
  • 930 nm Menlo YLMO Pulsed Laser
  • Neurogenetics
  • Neural Development and Plasticity

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Sam Tesfai
Sam Tesfai
Imaging Systems General Manager

Questions?
Need a Quote?

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Trans-Illumination Path Add-On for Rotating Bergamo Systems
Click to Enlarge
Trans-Illumination Add-On for Rotating Bergamo II Systems

Retrofit Options for Existing Multiphoton Systems

  • Upgrades to the Functionality of Existing Microscope Infrastructure and Components
  • Add-On Components to Increase Capabilities

Thorlabs' modular design enables our microscopes to continually evolve with experimental needs. Customers with previous model microscopes and product lines for multiphoton microscopy have the flexibility to update their infrustructure or add to their existing components as their microscopy needs change. See the expandable tables below for options available for each of our multiphoton system lines, and contact us for additional information about incorporating an upgrade or add-on into your system.

Please note that certain upgrades and add-ons require an on-site visit by one of our specialists for installation.

Bergamo II Compatible Upgrades and Add-Ons
Bergamo I (B-Scope) Compatible Upgrades and Add-Ons
Acerra (A-Scope) Compatible Upgrades and Add-Ons

Selected Publications Using Thorlabs' Imaging Systems

2023

 

Tsuji, M., Nishizuka, Y. & Emoto, K. (2023). Threat gates visual aversion via theta activity in Tachykinergic neurons. Nature Communications, 14(1), 3987. 

Villanueva, C. B., Stephensen, H. J. T., Mokso, R., Benraiss, A., Sporring, J., Goldman, S.A. (2023). Astrocytic engagement of the corticostriatal synaptic cleft is disrupted in a mouse model of Huntington's disease. Proceedings of the National Academy of Sciences, 120(24), e2210719120.

Holstein-Rønsbo, S., Gan, Y., Giannetto, M.J. et al. (2023). Glymphatic influx and clearance are accelerated by neurovascular coupling. Nature Neuroscience, 26, 1042–1053.

Serradj, N., Marino, F., Moreno-López, Y. et al. (2023). Task-specific modulation of corticospinal neuron activity during motor learning in mice. Nature Communications, 14(1), 2708.

Inayat, S., McAllister, B. B., Whishaw, I. Q., & Mohajerani, M. H. (2023). Hippocampal conjunctive and complementary CA1 populations relate sensory events to movement. Iscience, 26(4), 106481.

Untiet, V., Beinlich, F. R., Kusk, P., Kang, N., Ladrón-de-Guevara, A., Song, W., ... & Nedergaard, M. (2023). Astrocytic chloride is brain state dependent and modulates inhibitory neurotransmission in mice. Nature Communications, 14(1), 1871.

Anthoney, N., Tainton-Heap, L., Luong, H., Notaras, E., Zhao, Q., Perry, T., ... & van Swinderen, B. (2023). Experimentally induced active and quiet sleep engage non-overlapping transcriptomes in Drosophila. bioRxiv, 535331.

Boster, K. A., Cai, S., Ladrón-de-Guevara, A., Sun, J., Zheng, X., Du, T., ... & Kelley, D. H. (2023). Artificial intelligence velocimetry reveals in vivo flow rates, pressure gradients, and shear stresses in murine perivascular flows. Proceedings of the National Academy of Sciences, 120(14), e2217744120.

Scarpetta, V., Bodaleo, F., Salio, C., Agarwal, A., Sassoè-Pognetto, M., & Patrizi, A. (2023). Morphological and mitochondrial changes in murine choroid plexus epithelial cells during healthy aging. Fluids and Barriers of the CNS, 20(1), 1-16.

McDermott, K. D., Frechou, M. A., Jordan, J. T., Martin, S. S., & Gonçalves, J. T. (2023). Delayed formation of neural representations of space in aged mice. bioRxiv, 531021.

Matthews, M. D., Cook, E., Naguib, ,N., Wiesner, U. B., & Lewis, K. J. (2023). Intravital imaging of osteocyte integrin dynamics with locally injectable fluorescent nanoparticles. Bone, 174, 116830.

Tort-Colet, N., Resta, F., Montagni, E., Pavone, F., Allegra Mascaro, A. L., & Destexhe, A. (2023). Assessing brain state and anesthesia level with two-photon calcium signals. Scientific Reports, 13(1), 3183.

Esteves, I. M., Chang, H., Neumann, A. R., & McNaughton, B. L. (2023). Consolidation of cellular memory representations in superficial neocortex. Iscience, 26(2), 105970.

Quezada, A., Ward, C., Bader, E. R., Zolotavin, P., Altun, E., Hong, S., ... & Hébert, J. M. (2023). An in vivo platform for rebuilding functional neocortical tissue. Bioengineering, 10(2), 263.

Yang, J. Y., O’Connell, T. F., Hsu, W. M. M., Bauer, M. S., Dylla, K. V., Sharpee, T. O., & Hong, E. J. (2023). Restructuring of olfactory representations in the fly brain around odor relationships in natural sources. bioRxiv, 528627.

Carlton, A. J., Jeng, J. Y., Grandi, F. C., De Faveri, F., Ceriani, F., De Tomasi, L., ... & Marcotti, W. (2023). A critical period of prehearing spontaneous Ca2+ spiking is required for hair-bundle maintenance in inner hair cells. The EMBO Journal, 42(4), e112118.

Hamling, K. R., Zhu, Y., Auer, F., & Schoppik, D. (2023). Tilt In Place Microscopy (TIPM): a simple, low-cost solution to image neural responses to body rotations. Journal of Neuroscience, 43(6), 936-948.

Vestergaard, M., Carta, M., Güney, G., & Poulet, J. F. A. (2023). The cellular coding of temperature in the mammalian cortex. Nature, 614(7949), 725-731.

Contreras-López, R., Alatriste-León, H., Díaz-Hernández, E., Ramírez-Jarquín, J. O., & Tecuapetla, F. (2023). The deep cerebellar nuclei to striatum disynaptic connection contributes to skilled forelimb movement. Cell Reports, 42(1), 112000.

Mabuchi, Y., Cui, X., Xie, L., Kim, H., Jiang, T., & Yapici, N. (2023). GABA-mediated inhibition in visual feedback neurons fine-tunes Drosophila male courtship. bioRxiv, 525544.

Das, A., Margevicius, D., Borovicka, J., Icardi, J., Patel, D., Paquet, M. E., & Dana, H. (2023). Enhanced detection sensitivity of neuronal activity patterns using CaMPARI1 vs. CaMPARI2. Frontiers in Neuroscience, 16, 2291.

Lee, S., Lee, K., Choi, M., & Park, J. (2023). Implantable acousto-optic window for monitoring ultrasound-mediated neuromodulation in vivo. Neurophotonics, 9(3), 032203.

Zhuo, G. Y., Chen, M. C., Lin, T. Y., Lin, S. T., Chen, D. T. L., & Lee, C. W. S. (2023). Opioid-Modulated Receptor Localization and Erk1/2 Phosphorylation in Cells Coexpressing μ-Opioid and Nociceptin Receptors. International Journal of Molecular Sciences, 24(2), 1048.

Diaz-Cuadros, M., Miettinen, T. P., Skinner, O. S., Sheedy, D., Díaz-García, C. M., Gapon, S., ... & Pourquié, O. (2023). Metabolic regulation of species-specific developmental rates. Nature, 613(7944), 550-557.

Evrard, M., Mansuryan, T., Couderc, V., Désévédavy, F., Strutynski, C., Dussauze, M., ... & Smektala, F. (2023). Highly Nonlinear Multimode Tellurite Fibers: From Glass Synthesis to Practical Applications in Multiphoton Imaging. Advanced Photonics Research, 4(1), 2200213.

2022

 

Graham, R. T., Parrish, R. R., Alberio, L., Johnson, E. L., Owens, L., & Trevelyan, A. J. (2022). Optogenetic stimulation reveals a latent tipping point in cortical networks during ictogenesis. Brain, awac487.

Shiozaki, H. M., Wang, K., Lillvis, J. L., Xu, M., Dickson, B. J., & Stern, D. L. (2022). Neural coding of distinct motor patterns during Drosophila courtship song. bioRxiv, 520499.

Fisher, Y. E., Marquis, M., D’Alessandro, I., & Wilson, R. I. (2022). Dopamine promotes head direction plasticity during orienting movements. Nature, 612(7939), 316-322.

Wang, X., Delle, C., Asiminas, A., Akther, S., Vittani, M., Brxxxgger, P., ... & Hirase, H. (2022). Liver-secreted fluorescent blood plasma markers enable chronic imaging of the microcirculation. Cell Reports Methods, 2(10), 100302.

Lu, Y., Wei, X., Li, W., Wu, X., Chen, C., Li, G., ... & Gan, W. B. (2022). Large-volume and deep brain imaging in rabbits and monkeys using COMPACT two-photon microscopy. Scientific Reports, 12(1), 17736.

Huang, J. S., Kunkhyen, T., Rangel, A. N., Brechbill, T. R., Gregory, J. D., Winson-Bushby, E. D., ... & Cheetham, C. E. (2022). Immature olfactory sensory neurons provide behaviourally relevant sensory input to the olfactory bulb. Nature Communications, 13(1), 6194.

Govindaraju, I., Muraleedharan, M., Maidin, S., Chakraborty, I., Zhuo, G. Y., Mahato, K. K., & Mazumder, N. (2022). Investigation of the effect of gamma irradiation on the morphological structures of starch using advanced microscopic techniques. In Frontiers in Optics, JTu5A-74.

Hwang, F. J., Roth, R. H., Wu, Y. W., Sun, Y., Kwon, D. K., Liu, Y., & Ding, J. B. (2022). Motor learning selectively strengthens cortical and striatal synapses of motor engram neurons. Neuron, 110(17), 2790-2801.

Yue, Y., Ash, R. T., Boyle, N., Kinter, A., Li, Y., Zeng, C., & Lu, H. (2022). MeCP2 deficiency impairs motor cortical circuit flexibility associated with motor learning. Molecular Brain, 15(1), 1-12.

Liu, Z., Lu, X., Villette, V., Gou, Y., Colbert, K. L., Lai, S., Guan, S., ... & St-Pierre, F. (2022). Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy. Cell, 185(18), 3408-3425.

Aragon, M. J., Mok, A. T., Shea, J., Wang, M., Kim, H., Barkdull, N., ... & Yapici, N. (2022). Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle. Elife, 11, e69094.

Matheson, A. M., Lanz, A. J., Medina, A. M., Licata, A. M., Currier, T. A., Syed, M. H., & Nagel, K. I. (2022). A neural circuit for wind-guided olfactory navigation. Nature Communications, 13(1), 4613.

Chornyy, S., Borovicka, J. A., Patel, D., Shin, M. K., Vázquez-Rosa, E., Miller, E., ... & Dana, H. (2022). Longitudinal in vivo monitoring of axonal degeneration after brain injury. Cell Reports Methods3(5), 100481.

Sheng, W., Zhao, X., Huang, X., & Yang, Y. (2022). Real-time image processing toolbox for all-optical closed-loop control of neuronal activities. Frontiers in Cellular Neuroscience, 16, 917713.

Tadres, D., Shiozaki, H. M., Tastekin, I., Stern, D. L., & Louis, M. (2022). An essential experimental control for functional connectivity mapping with optogenetics. bioRxiv, 493610.

Koveal, D., Rosen, P. C., Meyer, D. J., Díaz-García, C. M., Wang, Y., Cai, L. H., ... & Yellen, G. (2022). A high-throughput multiparameter screen for accelerated development and optimization of soluble genetically encoded fluorescent biosensors. Nature Communications, 13(1), 2919.

Fukuda, M., Matsumura, T., Suda, T., & Hirase, H. (2022). Depth-targeted intracortical microstroke by two-photon photothrombosis in rodent brain. Neurophotonics, 9(2), 021910-021910.

Jackson, J. G., Krizman, E., Takano, H., Lee, M., Choi, G. H., Putt, M. E., & Robinson, M. B. (2022). Activation of Glutamate Transport Increases Arteriole Diameter in vivo: Implications for Neurovascular Coupling. Frontiers in Cellular Neuroscience, 16, 831061.

Du, T., Mestre, H., Kress, B. T., Liu, G., Sweeney, A. M., Samson, A. J., ... & Nedergaard, M. (2022). Cerebrospinal fluid is a significant fluid source for anoxic cerebral oedema. Brain, 145(2), 787-797.

Govindaraju, I., Zhuo, G. Y., Chakraborty, I., Melanthota, S. K., Mal, S. S., Sarmah, B., ... & Mazumder, N. (2022). Investigation of structural and physico-chemical properties of rice starch with varied amylose content: A combined microscopy, spectroscopy, and thermal study. Food Hydrocolloids, 122, 107093.

Choe, K., Hontani, Y., Wang, T., Hebert, E., Ouzounov, D. G., Lai, K., ... & Xu, C. (2022). Intravital three-photon microscopy allows visualization over the entire depth of mouse lymph nodes. Nature Immunology, 23(2), 330-340.

Spivak, Y. M., & Lemeshko, P. S. (2022). Multiphoton microscopy of mesoporous silicon. In Journal of Physics: Conference Series. 2227(1), 012013.

2021

 

Maset, A., Albanesi, M., di Soccio, A., Canova, M., Dal Maschio, M., & Lodovichi, C. (2021). Aberrant Patterns of Sensory-Evoked Activity in the Olfactory Bulb of LRRK2 Knockout Mice. Cells, 10(11), 3212.

Schmidt, E. R., Zhao, H. T., Park, J. M., Dipoppa, M., Monsalve-Mercado, M. M., Dahan, J. B., ... & Polleux, F. (2021). A human-specific modifier of cortical connectivity and circuit function. Nature, 599(7886), 640-644.

Hermans, L., Kaynak, M., Braun, J., Lobato Ríos, V., Chen, C. L., Günel, S., ... & Ramdya, P. (2021). Long-term imaging of the ventral nerve cord in behaving adult Drosophila. bioRxiv, 463778.

Carlsen, E. M. M., Falk, S., Skupio, U., Robin, L., Pagano Zottola, A. C., Marsicano, G., & Perrier, J. F. (2021). Spinal astroglial cannabinoid receptors control pathological tremor. Nature Neuroscience, 24(5), 658-666.

Moussa, N. O., Mansuryan, T., Hage, C. H., Fabert, M., Krupa, K., Tonello, A., ... & Couderc, V. (2021). Spatiotemporal beam self-cleaning for high-resolution nonlinear fluorescence imaging with multimode fiber. Scientific Reports, 11(1), 18240.

Eleftheriou, C. G., Corona, C., Khattak, S., Alam, N. M., Ivanova, E., Bianchimano, P., ... & Sagdullaev, B. T. (2021). Retinoschisin deficiency induces persistent aberrant waves of activity affecting neuroglial signaling in the retina. Journal of Neuroscience, 42(36), 6983-7000.

Han, J., Kim, S., Choi, P., Lee, S., Jo, Y., Kim, E., & Choi, M. (2021). Robust functional imaging of taste sensation with a Bessel beam. Biomedical Optics Express, 12(9), 5855-5864.

Favuzzi, E., Huang, S., Saldi, G. A., Binan, L., Ibrahim, L. A., Fernández-Otero, M., ... & Fishell, G. (2021). GABA-receptive microglia selectively sculpt developing inhibitory circuits. Cell, 184(15), 4048-4063.

Chornyy, S., Das, A., Borovicka, J. A., Patel, D., Chan, H. H., Hermann, J. K., ... & Dana, H. (2021). Cellular-resolution monitoring of ischemic stroke pathologies in the rat cortex. Biomedical Optics Express, 12(8), 4901-4919.

Malina, K. C. K., Tsivourakis, E., Kushinsky, D., Apelblat, D., Shtiglitz, S., Zohar, E., ... & Spiegel, I. (2021). NDNF interneurons in layer 1 gain-modulate whole cortical columns according to an animal’s behavioral state. Neuron, 109(13), 2150-2164.

Bruzzone, M., Chiarello, E., Albanesi, M., Miletto Petrazzini, M. E., Megighian, A., Lodovichi, C., & Dal Maschio, M. (2021). Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy. Scientific Reports, 11(1), 11048.

Mridha, Z., de Gee, J. W., Shi, Y., Alkashgari, R., Williams, J., Suminski, A., ... & McGinley, M. J. (2021). Graded recruitment of pupil-linked neuromodulation by parametric stimulation of the vagus nerve. Nature Communications, 12(1), 1539.

Ivanova, E., Corona, C., Eleftheriou, C. G., Bianchimano, P., & Sagdullaev, B. T. (2021). Retina-specific targeting of pericytes reveals structural diversity and enables control of capillary blood flow. Journal of Comparative Neurology, 529(6), 1121-1134.

Hung, C. W., Mazumder, N., Lin, D. J., Chen, W. L., Lin, S. T., Chan, M. C., & Zhuo, G. Y. (2021). Label-free characterization of collagen crosslinking in bone-engineered materials using nonlinear optical microscopy. Microscopy and Microanalysis, 27(3), 587-597.

Hontani, Y., Xia, F., & Xu, C. (2021). Multicolor three-photon fluorescence imaging with single-wavelength excitation deep in mouse brain. Science Advances, 7(12), eabf3531.

Resnik, J., & Polley, D. B. (2021). Cochlear neural degeneration disrupts hearing in background noise by increasing auditory cortex internal noise. Neuron, 109(6), 984-996.

Tainton-Heap, L. A., Kirszenblat, L. C., Notaras, E. T., Grabowska, M. J., Jeans, R., Feng, K., ... & van Swinderen, B. (2021). A paradoxical kind of sleep in Drosophila melanogaster. Current Biology, 31(3), 578-590.

Hortholary, T., Carrion, C., Chouzenoux, E., Pesquet, J. C., & Lefort, C. (2021). Multiplex-multiphoton microscopy and computational strategy for biomedical imaging. Microscopy Research and Technique, 84(7), 1553-1562.

Clayton, K. K., Williamson, R. S., Hancock, K. E., Tasaka, G. I., Mizrahi, A., Hackett, T. A., & Polley, D. B. (2021). Auditory corticothalamic neurons are recruited by motor preparatory inputs. Current Biology, 31(2), 310-321.

Esteves, I. M., Chang, H., Neumann, A. R., Sun, J., Mohajerani, M. H., & McNaughton, B. L. (2021). Spatial information encoding across multiple neocortical regions depends on an intact hippocampus. Journal of Neuroscience, 41(2), 307-319.

Schumacher, J. W., McCann, M. K., Maximov, K. J., & Fitzpatrick, D. (2021). Selective enhancement of neural coding in V1 underlies fine-discrimination learning in tree shrew. Current Biology, 32(15), 3245-3260.

2020

 

Keller, A. J., Dipoppa, M., Roth, M. M., Caudill, M. S., Ingrosso, A., Miller, K. D., & Scanziani, M. (2020). A disinhibitory circuit for contextual modulation in primary visual cortex. Neuron, 108(6), 1181-1193.

Lu, J., Behbahani, A. H., Hamburg, L., Westeinde, E. A., Dawson, P. M., Lyu, C., ... & Wilson, R. I. (2020). Transforming representations of movement from body-to world-centric space. Nature, 601(7891), 98-104.

Scholl, B., Thomas, C. I., Ryan, M. A., Kamasawa, N., & Fitzpatrick, D. (2020). Cortical response selectivity derives from strength in numbers of synapses. Nature, 590(7844), 111-114.

Robinson, N. T., Descamps, L. A., Russell, L. E., Buchholz, M. O., Bicknell, B. A., Antonov, G. K., ... & Häusser, M. (2020). Targeted activation of hippocampal place cells drives memory-guided spatial behavior. Cell, 183(6), 1586-1599.

Fan, J. L., Rivera, J. A., Sun, W., Peterson, J., Haeberle, H., Rubin, S., & Ji, N. (2020). High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics. Nature Communications, 11(1), 6020.

Okubo, T. S., Patella, P., D’Alessandro, I., & Wilson, R. I. (2020). A neural network for wind-guided compass navigation. Neuron, 107(5), 924-940.

Mazzarda, F., D'Elia, A., Massari, R., De Ninno, A., Bertani, F. R., Businaro, L., ... & Mammano, F. (2020). Organ-on-chip model shows that ATP release through connexin hemichannels drives spontaneous Ca 2+ signaling in non-sensory cells of the greater epithelial ridge in the developing cochlea. Lab on a Chip, 20(16), 3011-3023.

Bowen, Z., Winkowski, D. E., & Kanold, P. O. (2020). Functional organization of mouse primary auditory cortex in adult C57BL/6 and F1 (CBAxC57) mice. Scientific Reports, 10(1), 10905.

Feng, K., Sen, R., Minegishi, R., Dübbert, M., Bockemühl, T., Büschges, A., & Dickson, B. J. (2020). Distributed control of motor circuits for backward walking in Drosophila. Nature Communications, 11(1), 6166.

Keller, A. J., Roth, M. M., & Scanziani, M. (2020). Feedback generates a second receptive field in neurons of the visual cortex. Nature, 582(7813), 545-549.

Eschbach, C., Fushiki, A., Winding, M., Afonso, B., Andrade, I. V., Cocanougher, B. T., ... & Zlatic, M. (2020). Circuits for integrating learnt and innate valences in the fly brain. bioRxiv, 058339.

Montgomery, M. K., Kim, S. H., Dovas, A., Zhao, H. T., Goldberg, A. R., Xu, W., ... & Hillman, E. M. (2020). Glioma-induced alterations in neuronal activity and neurovascular coupling during disease progression. Cell Reports, 31(2), 107500.

Shiozaki, H. M., Ohta, K., & Kazama, H. (2020). A multi-regional network encoding heading and steering maneuvers in Drosophila. Neuron, 106(1), 126-141.

Mestre, H., Du, T., Sweeney, A. M., Liu, G., Samson, A. J., Peng, W., ... & Nedergaard, M. (2020). Cerebrospinal fluid influx drives acute ischemic tissue swelling. Science, 367(6483), eaax7171.

Jeng, J. Y., Ceriani, F., Hendry, A., Johnson, S. L., Yen, P., Simmons, D. D., ... & Marcotti, W. (2020). Hair cell maturation is differentially regulated along the tonotopic axis of the mammalian cochlea. The Journal of Physiology, 598(1), 151-170.

Kovacs-Oller, T., Ivanova, E., Bianchimano, P., & Sagdullaev, B. T. (2020). The pericyte connectome: spatial precision of neurovascular coupling is driven by selective connectivity maps of pericytes and endothelial cells and is disrupted in diabetes. Cell Discovery, 6(1), 39.

Oe, Y., Wang, X., Patriarchi, T., Konno, A., Ozawa, K., Yahagi, K., ... & Hirase, H. (2020). Distinct temporal integration of noradrenaline signaling by astrocytic second messengers during vigilance. Nature Communications, 11(1), 471.

2019

 

Romero, S., Hight, A. E., Clayton, K. K., Resnik, J., Williamson, R. S., Hancock, K. E., & Polley, D. B. (2019). Cellular and widefield imaging of sound frequency organization in primary and higher order fields of the mouse auditory cortex. Cerebral Cortex, 30(3), 1603-1622.

Nardin, C., Peres, C., Mazzarda, F., Ziraldo, G., Salvatore, A. M., & Mammano, F. (2019). Photosensitizer activation drives apoptosis by interorganellar Ca2+ transfer and superoxide production in bystander cancer cells. Cells, 8(10), 1175.

Scholl, B., Wilson, D. E., Jaepel, J., & Fitzpatrick, D. (2019). Functional logic of layer 2/3 inhibitory connectivity in the ferret visual cortex. Neuron, 104(3), 451-457.

Ceriani, F., Johnson, S. L., Sedlacek, M., Hendry, A., Kachar, B., Marcotti, W., & Mammano, F. (2019). Dynamic coupling of cochlear inner hair cell intrinsic Ca2+ action potentials to Ca2+ signaling of non-sensory cells. bioRxiv, 731851.

Brawek, B and Garaschuk O. (2019). Single-cell electroporation for measuring in vivo calcium dynamics in microglia. Microglia Methods and Protocols, 231-241.

Brawek, B., Olmedillas del Moral, M., & Garaschuk, O. (2019). In vivo visualization of microglia using tomato lectin. Microglia Methods and Protocols, 165-175.

Liang, Y. & Garaschuk O. (2019). Labeling microglia with genetically encoded calcium indicators. Microglia Methods and Protocols, 243–265.

Royzen, F., Williams, S., Fernandez, F. R., & White, J. A. (2019). Balanced synaptic currents underlie low-frequency oscillations in the subiculum. Hippocampus, 29(12), 1178-1189.

Rathore, A. P. S., Mantri, C. K., Aman, S. A. B., Syenina, A., Ooi, J., Jagaraj, C. J., ... & St. John, A. L. (2019). Dengue virus-elicited tryptase induces endothelial permeability and shock. Journal of Clinical Investiagtion, 129(10), 4180-4193.

Stringer, C., Pachitariu, M., Steinmetz, N., Carandini, M., & Harris, K. D. (2019). High-dimensional geometry of population responses in visual cortex. Nature, 571(7765), 361–365.

Ziraldo, G., Buratto, D., Kuang, Y., Xu, L., Carrer, A., Nardin, C., ... & Mammano, F. (2019). A human-derived monoclonal antibody targeting extracellular connexin domain selectively modulates hemichannel function. Frontiers in Physiology, 10, 392.

Díaz-García, C. M., Lahmann, C., Martínez-François, J. R., Li, B., Koveal, D., Nathwani N., ... & Yellen, G. (2019). Quantitative in vivo imaging of neuronal glucose concentrations with a genetically encoded fluorescence lifetime sensor. Journal of Neuroscience Research, 97(8), 946–960.

Philip, V., Newton, D., Oh, H., Collins, S., Bercik, P., & Sibille, E. (2019). The Effect of Gut Microbiota on Glutamatergic/GABAergic Gene Expression in Adult Mice. Biological Psychiatry, 85, S127–S128.

Bowen, Z., Winkowski, D. E., Seshadri, S., Plenz, D., & Kanold, P. O. (2019). Neuronal avalanches in input and associative layers of auditory cortex. Frontiers in Systems Neuroscience, 13, 45.

Burgold, J., Schulz-Trieglaff, E. K., Voelkl, K., Gutiérrez-Ángel, S., Bader, J. M., Hosp, F., ... & Dudanova, I. (2019). Cortical circuit alterations precede motor impairments in Huntington’s disease mice. Scientific Reports, 9(1), 6634.

Matovic, S., Ichiyama, A., Igarashi, H., Salter, E. W., Wang, X. F., Henry, M., ... & Inoue, W. (2019). Stress-induced neuronal hypertrophy decreases the intrinsic excitability in stress habituation. bioRxiv, 593665.

Weissenberger, Y., King, A. J., & Dahmen, J. C. (2019). Decoding mouse behavior to explain single-trial decisions and their relationship with neural activity. bioRxiv, 567479.

Ceriani, F., Hendry, A., Jeng, J. Y., Johnson, S. L., Stephani, F., Olt, J., ... & Marcotti, W. (2019). Coordinated calcium signalling in cochlear sensory and non-sensory cells refines afferent innervation of outer hair cells. The EMBO Journal, 38(9), e99839.

Lee, K. S., Vandemark, K., Mezey, D., Shultz, N., & Fitzpatrick, D. (2019). Functional synaptic architecture of callosal inputs in mouse primary visual cortex. Neuron, 101(3), 421-428.

2018

 

Marvin, J. S., Scholl, B., Wilson, D. E., Podgorski, K., Kazemipour, A., Müller, J. A., ... & Looger, L. L. (2018). Stability, affinity, and chromatic variants of the glutamate sensor iGluSnFR. Nature Methods, 15(11), 936–939.

Moeyaert, B., Holt, G., Madangopal, R., Perez-Alvarez, A., Fearey, B. C., Trojanowski, N. F., ... & Schreiter, E. R. (2018). Improved methods for marking active neuron populations. Nature Communications, 9(1), 440.

Chen, C. L., Hermans, L., Viswanathan, M. C., Fortun, D., Aymanns, F., Unser, M., ... & Ramdya, P. (2018). Imaging neural activity in the ventral nerve cord of behaving adult Drosophila. Nature Communications9(1), 4390.

Corns, L. F., Johnson, S. L., Roberts, T., Ranatunga, K. M., Hendry, A., Ceriani, F., ... & Marcotti, W. (2018). Mechanotransduction is required for establishing and maintaining mature inner hair cells and regulating efferent innervation. Nature Communications, 9(1), 4015.

Saleem, A. B., Diamanti, E. M., Fournier, J., Harris, K. D., & Carandini, M. (2018). Coherent encoding of subjective spatial position in visual cortex and hippocampus. Nature, 562(7725), 124-127.

Dipoppa, M., Ranson, A., Krumin, M., Pachitariu, M., Carandini, M., & Harris, K. D. (2018). Vision and locomotion shape the interactions between neuron types in mouse visual cortex. Neuron, 98(3), 602-615.

Gillet, S. N., Kato, H. K., Justen, M. A., Lai, M., & Isaacson, J. S. (2018). Fear learning regulates cortical sensory representations by suppressing habituation. Frontiers in Neural Circuits, 11, 112.

2017

 

Scholl, B., Wilson, D. E., & Fitzpatrick, D. (2017). Local order within global disorder: synaptic architecture of visual spaceNeuron, 96(5), 1127-1138.

Klapoetke, N. C., Nern, A., Peek, M. Y., Rogers, E. M., Breads, P., Rubin, G. M., ... & Card, G. M. (2017). Ultra-selective looming detection from radial motion opponencyNature Neuroscience, 551(7679), 237-241.

Kato, H. K., Asinof, S. K., & Isaacson, J. S. (2017). Network-level control of frequency tuning in auditory cortexNeuron, 95(2), 412-423.

Lu, R., Sun, W., Liang, Y., Kerlin, A., Bierfeld, J., Seelig, J.D., W... & Ji, N. (2017). Video-rate volumetric functional imaging of the brain at synaptic resolutionNature Neuroscience, 20(4), 620-628.

2016

 

Mongeon, R., Venkatachalam, V., & Yellen, G. (2016). Cytosolic NADH-NAD+ redox visualized in brain slices by two-photon fluorescence lifetime biosensor imaging.Antioxid Redox Signal, 25(10), 553-563.

Pachitariu, M., Stringer, C., Schröder, S., Dipoppa, M., Rossi, L. F., Carandini, M., & Harris, K. D. (2016). Suite2p: beyond 10,000 neurons with standard two-photon microscopybioRxiv, 061507.

Rose, T., Jaepel, J., Hübener, M., & Bonhoeffer, T. (2016). Cell-specific restoration of stimulus preference after monocular deprivation in the visual cortexScience, 352(6291), 1319–1322.

Strobl, M. J., Freeman, D., Patel, J., Poulsen, R., Wendler, C. C., Rivkees, S. A., & Coleman, J. E. (2016). Opposing effects of maternal hypo-and hyperthyroidism on the stability of thalamocortical synapses in the visual cortex of adult offspring.Cerebral Cortex, 27(5), 3015-3027.

Lee, K. S., Huang, X., & Fitzpatrick, D. (2016). Topology of ON and OFF inputs in visual cortex enables an invariant columnar architectureNature, 533(7601), 90-94.

Monai, H., Ohkura, M., Tanaka, M., Oe, Y., Konno, A., Hirai, H., ... & Hirase, H. (2016). Calcium imaginq reveals glial involvement in transcranial direct current stimulation-induced plasticity in mouse brain. Nature Communications7(1), 11100.

Ganmor, E., Krumin, M., Rossi, L. F., Carandini, M., & Simoncelli, E. P. (2016). Direct estimation of firing rates from calcium imaging data. arXiv, 1601.00364.

2015

 

Roth, M. M., Dahmen, J.C., Muir, D. R., Imhof, F., Martini, F. J., & Hofer, S. B. (2015). Thalamic nuclei convey diverse contextual information to layer 1 of visual cortexNature Neuroscience, 19(2), 299-307.

Barnstedt, O., Keating, P., Weissenberger, Y., King, A. J., & Dahmen, J. C. (2015). Functional microarchitecture of the mouse dorsal inferior colliculus revealed through in vivo two-photon calcium imaging.. Journal of Neuroscience, 35(31), 10927-10939.

Chen, S. X., Kim, A. N., Peters, A. J., & Komiyama, T. (2015). Subtype-specific plasticity of inhibitory circuits in motor cortex during motor learningNature Neuroscience, 18(8), 1109-1115.

Jia, Y., Zhang, S., Miao, L., Wang, J., Jin, Z., Gu, B., ... & Li, Z. (2015). Activation of platelet protease-activated receptor-1 induces epithelial-mesenchymal transition and chemotaxis of colon cancer cell line SW620Oncology Reports, 33(6), 2681-2688.

Lu, W., Tang, Y., Zhang, Z., Zhang, X., Yao, Y., Fu, C., ... & Ma, G. (2015). Inhibiting the mobilization of Ly6Chigh monocytes after acute myocardial infarction enhances the efficiency of mesenchymal stromal cell transplantation and curbs myocardial remodeling.American Journal of Translational Research, 7(3), 587-597.

Boyd, A. M., Kato, H. K., Komiyama, T., & Isaacson, J. S. (2015). Broadcasting of cortical activity to the olfactory bulbCell Reports, 10(7), 1032-1039.

Cossell, L., Iacaruso, M. F., Muir, D. R., Houlton, R., Sader, E. N., Ko, H., H... & Mrsic-Flogel, T. D. (2015). Functional organization of excitatory synaptic strength in primary visual cortexNature, 518(7539), 399-403.

2014

 

Partridge, J. G., Lewin, A. E., Yasko, J. R., & Vicini, S. (2014). Contrasting actions of group I metabotropic glutamate receptors in distinct mouse striatal neuronesJournal of Physiology, 592(13), 2721-2733.

Peters, A. J., Chen, S, X., Komiyama, T. (2014). Emergence of reproducible spatiotemporal activity during motor learningNature, 510(7504), 263-267.

Ehmke, T., Nitzsche, T. H., Knebl, A., & Heisterkamp, A. (2014). Molecular orientation sensitive second harmonic microscopy by radially and azimuthally polarized lightBiomedical Optics Express, 5(7), 2231-46.

Liu, J., Wu, N., Ma, L., Liu, M., Liu, G., Zhang, Y., & Lin, X. (2014). Oleanolic acid suppresses aerobic glycolysis in cancer cells by switching pyruvate kinase type M isoformsPLoS One, 9(3), e91606.

Palmer, L. M., Shai, A. S., Reeve, J.E., Anderson, H. L., Paulsen, O., & Larkum, M. E. (2014). NMDA spikes enhance action potential generation during sensory inputNature Neuroscience17(3), 383-390.

Cai, F., Yu, J., Qian, J., Wang, Y., Chen, Z., Huang, J., ... & He, S. (2014). Use of tunable second-harmonic signal from KNbO3 nanoneedles to find optimal wavelength for deep-tissue imagingLaser & Photonics Reviews, 8(6), 865-874.

2013

 

Kato, H. K., Gillet, S. N., Peters, A. J., Isaacson, J. S., & Komiyama, T. (2013). Parvalbumin-expressing interneurons linearly control olfactory bulb outputNeuron, 80(5), 1218-1231.

Takata, N., Nagai, T., Ozawa, K., Oe, Y., Mikoshiba, K., & Hirase, H. (2013). Cerebral blood flow modulation by Basal forebrain or whisker stimulation can occur independently of large cytosolic Ca2+ signaling in astrocytesPLoS One, 8(6), e66525.

The full source code for ThorImage®LS is available for owners of a Bergamo®, Cerna® Confocal, Veneto®, or confocal microscope. Click here to receive your copy.

ThorImageLS Brochure

ThorImage®LS Software

ThorImageLS is an open-source image acquisition program that controls Thorlabs' microscopes, as well as supplementary external hardware. From prepared-slice multiphoton Z-stacks to simultaneous in vivo photoactivation and imaging, ThorImageLS provides an integrated, modular workspace tailored to the individual needs of the scientist. Its workflow-oriented interface supports single image, Z-stacks, time series, and image streaming acquisition, visualization, and analysis. See the video to the lower right for a real-time view of data acquisition and analysis with ThorImageLS.

ThorImageLS is included with a Thorlabs microscope system purchase and is open source, allowing full customization of software features and performance. ThorImageLS also includes Thorlabs’ customer support and regular software updates to continually meet the imaging demands of the scientific community.

For additional details, see the full web presentation.

Advanced Software Functionality

  • Multi-Column Customizable Workspace
  • Image Acquisition Synced with Hardware Inputs and Timing Events
  • Live Image Correction and ROI Analysis
  • Independent Galvo-Galvo and Galvo-Resonant Scan Areas and Geometries
  • Tiling for High-Resolution Large-Area Imaging
  • Independent Primary and Secondary Z-Axis Control for Fast Deep-Tissue Scans
  • Automated Image Capture with Scripts
    • Compatible with ImageJ Macros
  • Multi-User Settings Saved for Shared Workstations
  • Individual Colors for Detection Channels Enable Simple Visual Analysis

Seamless Integration with Experiments

  • Simultaneous Multi-Point Photoactivation and Imaging with Spatial Light Modulator
  • Fast Z Volume Acquisition with PFM450E or Third-Party Objective Scanners
  • Electrophysiology Signaling
  • Wavelength Switching with Tiberius® Laser or Coherent Chameleon Lasers
  • Pockels Cell ROI Masking
  • Power Ramped with Depth to Minimize Damage and Maximize Signal-to-Noise

Features of ThorImage®LS

Brochures

The buttons below link to PDFs of printable materials for Bergamo® II microscopes.

Bergamo II Brochure ThorImageLS Brochure Tiberius Brochure Download

Laser Scanning
Scan Path Wavelength Range 450 - 1100 nm, 680 - 1300 nm, or 800 - 1800 nm
Scan Paths Resonant-Galvo-Galvo Scanner, Galvo-Resonant Scanners,
Galvo-Galvo Scanners, or Spatial Light Modulator;
Single or Dual Scan Paths
Scan Speed 8 kHz Resonant-Galvo-Galvo
or Galvo-Resonant
2 fps at 4096 x 4096 Pixels
30 fps at 512 x 512 Pixels
400 fps at 512 x 32 Pixels
12 kHz Resonant-Galvo-Galvo
or Galvo-Resonant
4.4 fps at 2048 x 2048 Pixels
45 fps at 512 x 512 Pixels
600 fps at 512 x 32 Pixels
Galvo-Galvo 3 fps at 512 x 512 Pixels
48 fps at 512 x 32 Pixels
70 fps at 32 x 32 Pixels
Pixel Dwell Time: 0.4 to 20 µs
Galvo-Galvo Scan Modes Imaging: Line, Polyline, Square, or Rectangle
Non-Imaging: Circle, Ellipse, Polygon, or Point
Field of View 20 mm Diagonal Square (Max) at the Intermediate Image Plane
[12 mm Diagonal Square (Max) for 12 kHz Scanner]
Scan Zoom 1X to 16X (Continuously Variable)
Scan Resolution Up to 2048 x 2048 Pixels (Bi-Directional) [Up to 1168 x 1168 Pixels for 12 kHz Scanners]
Up to 4096 x 4096 Pixels (Unidirectional) [Up to 2336 x 2336 Pixels for 12 kHz Scanners]
Compatible Objective Threadings M34 x 1.0, M32 x 0.75, M25 x 0.75, and RMS
Multiphoton Signal Detection
Epi-Detection Up to Four Ultrasensitive GaAsP PMTs, Cooled or Non-Cooled
Forward-Direction Detection Two Ultrasensitive GaAsP PMTs
Maximum of Four PMTs Controlled by the Software at a Given Time
Collection Optics 8°, 10°, or 14° Collection Angle
(Angles Quoted When Using an Objective with a 20 mm Entrance Pupil)
Easy-to-Exchange Emission Filters and Dichroic Mirrors
Confocal Imaging
Motorized Pinhole Wheel with 16 Round Pinholes from Ø25 µm to Ø2 mm
Two to Four Laser Lines (488 nm Standard; Other Options Range from 405 nm to 660 nm)
Standard Multialkali or High-Sensitivity GaAsP PMTs
Easy-to-Exchange Emission Filters and Dichroic Mirrors
Widefield Viewing
Manual or Motorized Switching Between Scanning and Widefield Modes
Illumination Provided via LED or Liquid Light Guide
C-Mount Threads for Scientific Cameras
Transmitted Light Imaging
Differential Interference Contrast (DIC) or Dodt Gradient Contrast
Widefield or Laser Scanned
Illumination Provided by Visible and/or NIR LEDs
Compatible with Air or Oil Immersion Condensers
Three-Photon Imaging
Scan Optics for 800 - 1800 nm Range
Achieve Reduced Background Scatter for Greater Sensitivity in Deep Tissue Imaging
Volume Imaging Using Bessel Beams
3D Volumetric Functional Imaging at Video Frame Rates
Enhanced Temporal Resolution for Studying Internal Systems at Cellular Lateral Resolution In Vivo
Translation
Microscope Body Rotation
(Rotating Bodies Only)
0° to 90° or -45° to +45° Around Objective Focus
0.1° Encoder Resolution
Coarse Elevator Base Z
(Rotating Bodies Only)
5" (127 mm) Total Travel; 1 µm Encoder Resolution
Fine Microscope Body X and Y 2" (50.8 mm) Total Travel; 0.5 µm Encoder Resolution
Fine Microscope Arm Z  1" (25.4 mm) Total Travel; 0.1 µm Encoder Resolution
Fine Objective Z
(Piezo Objective Scanner)
Open Loop: 600 µm ± 10% Travel Range; 1 nm Resolution
Closed Loop: 450 µm Travel Range; 3 nm Resolution

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

To schedule an in-person or virtual demo appointment, please email ImagingSales@thorlabs.com.

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Try Our Microscopes In Person or Virtually

Thorlabs' sales engineers and field service staff are based out of nine offices across four continents. We look forward to helping you determine the best imaging system to meet your specific experimental needs. Our customers are attempting to solve biology's most important problems; these endeavors require matching systems that drive industry standards for ease of use, reliability, and raw capability.

Thorlabs' worldwide network allows us to operate demo rooms in a number of locations where you can see our systems in action. We welcome the opportunity to work with you in person or virtually. A demo can be scheduled at any of our showrooms or virtually by contacting ImagingSales@thorlabs.com.

Customer Support Sites
(Click Each Location for More Details)

Demo Rooms and Customer Support Sites
(Click Each Location for More Details)


Posted Comments:
gaiqing Wang  (posted 2020-05-07 11:13:33.577)
I am looking for a cheap way to do confocal imaging in vivo. Is this Bergamo II Series Multiphoton Microscope my best option? Can you send me a quote?
YLohia  (posted 2020-05-07 09:45:11.0)
Thank you for contacting Thorlabs. We will reach out to you directly to discuss your requirements.
jfpena  (posted 2016-12-19 18:15:55.003)
I am looking for a cheap way to do confocal imaging in vivo. Is this Bergamo II Series Multiphoton Microscope my best option? Can you send me a quote?
tfrisch  (posted 2016-12-22 11:44:31.0)
Hello, thank you for contacting Thorlabs. A member of our Imaging Team will reach out to you directly to discuss this system and your application.
birech  (posted 2016-11-17 06:33:49.463)
I asked for a price quote for this product, Bergamo II Series Multiphoton Microscopes three days ago. I am working at the University of Nairobi in Kenya and would wish to order one. Regards, Birech
tfrisch  (posted 2016-11-17 06:56:23.0)
Hello, thank you for contacting Thorlabs. I have forwarded this request to our Imaging Sales Team. I apologize for the delay.
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Last Edited: Nov 08, 2013 Author: Dan Daranciang