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Microscope Objective, Tube, and Scan Lens Tutorials
Browse Our Selection of Objective, Scan, and Tube Lenses
Note: While the diagrams above show typical markings, they serve only as examples. The format and content of the engraved specifications will vary between objectives and manufacturers.
Types of Objectives
Thorlabs offers several types of objectives to meet a variety of experimental needs. This guide describes the features and benefits of each type of objective.
Dry, Immersion, and Dipping Objectives
Plan Achromat and Plan Apochromat Objectives
Plan Fluorite Objectives
Super Apochromat Objectives
Glossary of Terms
M = L / F .
The total magnification of the system is the magnification of the objective multiplied by the magnification of the eyepiece or camera tube. The specified magnification on the microscope objective housing is accurate as long as the objective is used with a compatible tube lens focal length.
Numerical Aperture (NA)
NA = ni × sinθa
where θa is the maximum 1/2 acceptance angle of the objective, and ni is the index of refraction of the immersion medium. This medium is typically air, but may also be water, oil, or other substances.
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This graph shows the effect of a cover slip on image quality at 632.8 nm.
FN = Field of View Diameter × Magnification
Coverslip Correction and Correction Collar (Ring)
The graph to the right shows the magnitude of spherical aberration versus the thickness of the coverslip used, for 632.8 nm light. For the typical coverslip thickness of 0.17 mm, the spherical aberration caused by the coverslip does not exceed the diffraction-limited aberration for objectives with NA up to 0.40.
When viewing an image with a camera, the system magnification is the product of the objective and camera tube magnifications. When viewing an image with trinoculars, the system magnification is the product of the objective and eyepiece magnifications.
Magnification and Sample Area Calculations
The magnification of a system is the multiplicative product of the magnification of each optical element in the system. Optical elements that produce magnification include objectives, camera tubes, and trinocular eyepieces, as shown in the drawing to the right. It is important to note that the magnification quoted in these products' specifications is usually only valid when all optical elements are made by the same manufacturer. If this is not the case, then the magnification of the system can still be calculated, but an effective objective magnification should be calculated first, as described below.
To adapt the examples shown here to your own microscope, please use our Magnification and FOV Calculator, which is available for download by clicking on the red button above. Note the calculator is an Excel spreadsheet that uses macros. In order to use the calculator, macros must be enabled. To enable macros, click the "Enable Content" button in the yellow message bar upon opening the file.
Example 1: Camera Magnification
Example 2: Trinocular Magnification
Using an Objective with a Microscope from a Different Manufacturer
Magnification is not a fundamental value: it is a derived value, calculated by assuming a specific tube lens focal length. Each microscope manufacturer has adopted a different focal length for their tube lens, as shown by the table to the right. Hence, when combining optical elements from different manufacturers, it is necessary to calculate an effective magnification for the objective, which is then used to calculate the magnification of the system.
The effective magnification of an objective is given by Equation 1:
Here, the Design Magnification is the magnification printed on the objective, fTube Lens in Microscope is the focal length of the tube lens in the microscope you are using, and fDesign Tube Lens of Objective is the tube lens focal length that the objective manufacturer used to calculate the Design Magnification. These focal lengths are given by the table to the right.
Note that Leica, Mitutoyo, Nikon, and Thorlabs use the same tube lens focal length; if combining elements from any of these manufacturers, no conversion is needed. Once the effective objective magnification is calculated, the magnification of the system can be calculated as before.
Example 3: Trinocular Magnification (Different Manufacturers)
Following Equation 1 and the table to the right, we calculate the effective magnification of an Olympus objective in a Nikon microscope:
The effective magnification of the Olympus objective is 22.2X and the trinoculars have 10X eyepieces, so the image at the eyepieces has 22.2X × 10X = 222X magnification.
Sample Area When Imaged on a Camera
When imaging a sample with a camera, the dimensions of the sample area are determined by the dimensions of the camera sensor and the system magnification, as shown by Equation 2.
The camera sensor dimensions can be obtained from the manufacturer, while the system magnification is the multiplicative product of the objective magnification and the camera tube magnification (see Example 1). If needed, the objective magnification can be adjusted as shown in Example 3.
As the magnification increases, the resolution improves, but the field of view also decreases. The dependence of the field of view on magnification is shown in the schematic to the right.
Example 4: Sample Area
Sample Area Examples
The images of a mouse kidney below were all acquired using the same objective and the same camera. However, the camera tubes used were different. Read from left to right, they demonstrate that decreasing the camera tube magnification enlarges the field of view at the expense of the size of the details in the image.
Scan lenses are used in a variety of laser imaging systems, including confocal laser scanning microscopy, optical coherence tomography (OCT), and multiphoton imaging systems. In these applications, a laser beam incident on the back aperture (entrance pupil) of the lens is scanned through a range of angles. This translates the position of the spot formed in the image plane across the lens' field of view. In the case of non-telecentric lenses, this approach to scanning the focal spot through the image plane would introduce severe aberrations that would significantly degrade the quality of the resulting image. Telecentric scan lenses are designed to create a uniform spot size in the image plane at every scan position, which allows a high-quality image of the sample to be formed. Plots showing the spot size and scan position as a function of scan angle are included in the Specs tab.
In general, laser scanning microscopy systems pair a scan lens with a tube lens to create an infinity-corrected optical system. However, most OCT systems are designed to use the scan lens without a tube lens. The CLS-SL, SL50-CLS2, SL50-2P2, and SL50-3P lenses were optimized for use in Thorlabs' confocal laser scanning and multiphoton microscopy systems, and the LSM family of lenses were optimized to be used in OCT imaging systems. A brief discussion of scanning systems implemented with and without tube lenses follows.
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The tube and scan lens schematic above shows the lens spacing for the SL50-2P2 scan lens used with a 200 mm focal length telecentric tube lens. Note that for the SL50-CLS2, the entrance pupil at the scan plane is a maximum of Ø4 mm.
Scan Lenses Implemented in General Laser Scanning Microscopy Applications
The image to the right shows the proper spacing of the scan and tube lenses for laser scanning microscopy. The scanning mirror, which is located at the left of the image at the scan plane, directs the laser beam through the scan lens. The angle at which the laser beam is incident on the scan lens determines the position of the focal spot in the intermediate image plane, which is located between the scan lens and the ITL200 tube lens. The tube lens is positioned so that it collects and collimates the light (the focus is at infinity). The collimated light is collected by the objective, which brings it to a focus on the sample plane. Light scattered or emitted from the sample plane is collected by the objective and directed to a detector. The image below and to the left shows a CLS-SL scan lens paired with a tube lens; clicking on the image shows the correct spacing for using the CLS-SL with the ITL200 tube lens.
An attractive feature of this optical system design is the collimated light that is produced as a result of pairing the scan lens with the tube lens. With the light from the tube lens focusing at infinity, it is possible to move the position of the objective with respect to the tube lens without impacting the image quality at the sample plane. This imparts considerable flexibility to the design of the optical system. If no tube lens were used, the scan lens would also function as the objective and the intermediate image plane would become the sample plane. It would not be possible to move the image plane much with respect to the scan lens while maintaining image quality.
The image below and to the right shows the relationship between the scan distance and the objective distance. In a perfect 4f optical system (using the CLS-SL as an example), d1 = 52 mm (minimum scan distance) and d2 = f2. However, in many practical cases the system is slightly deviated from this perfect alignment. For instance, in many commercial microscopes, the objective distance (d2) is not the same as the focal length (f2), so there may be a need to adjust distances. The figure below and to the right shows the scan and objective distance moved by some small distance δ1 and δ2, respectively. The relationship between these values is δd1 = -δd2*(f1/f2)2.
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CLS-SL Tube Lens integration with the ITL200 and an objective in a laser scanning system.
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A diagram showing lens placement for a general laser scanning system.
Scan Lenses Implemented in OCT
When designing an imaging system that uses an LSM scan lens in an OCT configuration, it is important to accommodate the design wavelength, parfocal distance, scanning distance, entrance pupil, and scan angle specifications in order to maximize the image quality (see the Specs tab for scan lens specifications and definitions). In general, the larger the input beam diameter, the smaller the focused spot size. However, due to the effects of vignetting and/or increased aberrations, the range of scan angles decreases as the diameter of the beam increases. Beams smaller than the entrance pupil specification will result in spot sizes larger than those specified in the Specs tab, and beams with larger diameters will be clipped.
For imaging systems with a single galvo mirror the center of the scan lens' entrance pupil is coincident with the pivot point of the galvo mirror. When a single galvo mirror is used, the scanning distance is measured from the mounting surface of the lens to the pivot point of the mirror. This is shown in the image at bottom-left.
When one galvo mirror is used, the entrance pupil is located
at the pivot point of the mirror.
When two galvo mirrors are used, the entrance pupil is located between the mirrors.