Insights into Best Lab Practices

Scroll down to read about things to consider when building integrating spheres into setups and analyzing data results.

• Ultraviolet and Blue Fluorescence Emitted by Integrating Spheres
• Sample Substitution Errors

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Ultraviolet and Blue Fluorescence Emitted by Integrating Spheres

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Figure 1: Typical yields at each wavelength are around four orders of magnitude lower than the excitation wavelength. [4]

The spectral fluorescence yield relates the intensity of the fluorescence emitted within the integrating sphere with the intensity of the excitation wavelength. The yield is calculated by dividing the wavelength-dependent, total fluorescence excited over the entire interior surface of the sphere by the intensity of the light excitation.

Data were kindly provided by Dr. Ping-Shine Shaw, Physics Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.

A material of choice for coating the light-diffusing cavities of integrating spheres is polytetrafluoroethylene (PTFE). This material, which is white in appearance, is favored for reasons including its high, flat reflectance over a wide range of wavelengths (see the Specs tab for details) and chemical inertness.

However, it should be noted that integrating spheres coated with both PTFE and barium sulfate, which is an alternative coating with lower reflectance, emit low levels of ultraviolet (UV) and blue fluorescence when irradiated by UV light. [1-3]

Hydrocarbons in the PTFE Fluoresce
It is not the PTFE that fluoresces. The sources of the UV and blue fluorescence are hydrocarbons in the PTFE. Low levels of hydrocarbon impurities are present in the raw coating material, and pollution sources deposit additional hydrocarbon contaminants in the PTFE material of the integrating sphere during its use and storage. [1]

Fluorescence Wavelength Bands and Strength
Researchers at the National Institute of Standards and Technology (NIST) have investigated the fluorescence excited by illuminating PTFE-coated integrating spheres. The total fluorescence output by the integrating sphere was measured with respect to fluorescence wavelength and excitation wavelength. The maximum fluorescence was approximately four orders of magnitude lower than the intensity of the exciting radiation.

The UV and blue fluorescence from PTFE is primarily excited by incident wavelengths in a 200 nm to 300 nm absorption band. The fluorescence is emitted in the 250 nm to 400 nm wavelength range, as shown by Figure 1. These data indicate that increasing the excitation wavelength decreases the fluorescence emitted at lower wavelengths and changes the shape of the fluorescence spectrum.

As the levels of hydrocarbon contaminants in the PFTE increase, fluorescence increases. A related effect is a decrease of the light output by the integrating sphere over the absorption band wavelengths, due to more light from this spectral region being absorbed. [1, 3]

Impact on Applications
The UV and blue fluorescence from the PTFE has negligible effect on many applications, since the intensity of the fluorescence is low and primarily excited by incident wavelengths <300 nm. Applications sensitive to this fluorescence include long-term measurements of UV radiation throughput, UV source calibration, establishing UV reflectance standards, and performing some UV remote sensing tasks. [1]

Minimizing Fluorescence Effects
Minimizing and stabilizing the fluorescence levels requires isolating the integrating sphere from all sources of hydrocarbons, including gasoline- and diesel-burning engine exhaust and organic solvents, such as naphthalene and toluene. It should be noted that, while hydrocarbon contamination can be minimized and reduced, it cannot be eliminated. [1]

Since the history of each integrating sphere's exposure to hydrocarbon contaminants is unique, it is not possible to predict the response of a particular sphere to incident radiation. When an application is negatively impacted by the fluorescence, calibration of the integrating sphere is recommended. A calibration procedure described in [4] requires a light source with a well-known spectrum that extends across the wavelength region of interest, such as a deuterium lamp or synchrotron radiation, a monochromator, a detector, and the integrating sphere.

References
[1] Ping-Shine Shaw, Zhigang Li, Uwe Arp, and Keith R. Lykke, "Ultraviolet characterization of integrating spheres," Appl.Opt. 46, 5119-5128 (2007).
[2] Jan Valenta, "Photoluminescence of the integrating sphere walls, its influence on the absolute quantum yield measurements and correction methods," AIP Advances 8, 102123 (2018).
[3] Robert D. Saunders and William R. Ott, "Spectral irradiance measurements: effect of UV-produced fluorescence in integrating spheres," Appl. Opt. 15, 827-828 (1976).
[4] Ping-Shine Shaw, Uwe Arp, and Keith R. Lykke, "Measurement of the ultraviolet-induced fluorescence yield from integrating spheres," Metrologia 46, S191 - S196 (2009).

Date of Last Edit: Dec. 4, 2019

Sample Substitution Errors

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Figure 2: Measuring diffuse sample transmittance and reflectance as shown above can result in a distorted sample spectrum due to sample substitution error. The problem is that the reflectivity over the sample area is different during the reference and sample measurements.

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Figure 3: The above configuration is not susceptible to sample substitution error, since the interior of the sphere is the same for reference and sample measurements. During the reference measurement the light travels along (R), and no light is incident along (S). The opposite is true when a sample measurement is made.

Absolute transmittance and absolute diffuse reflectance spectra of optical samples can be found using integrating spheres. These spectra are found by performing spectral measurements of both the sample of interest and a reference.

Measurement of a reference is needed since this provides the spectrum of the illuminating light source. Obtaining the reference scan allows the spectrum of the light source to be subtracted from the sample measurement.

The light source reference measurement is made with no sample in place for transmittance data and with a highly reflective white standard reference sample in place for reflectance measurements.

Sample substitution errors incurred while acquiring the sample and reference measurement sets can negatively effect the accuracy of the corrected sample spectrum, unless the chosen experimental technique is immune to these errors.

Conditions Leading to Sample Substitution Errors
An integrating sphere's optical performance depends on the reflectance at each point on its entire inner surface. Often, a section of the sphere's inner wall is replaced by the sample when its transmittance and diffuse reflectance spectra are measured (Figure 2). However, modifying a section of the inner wall alters the performance of the integrating sphere.

Sample substitution errors are a concern when the measurement procedure involves physically changing one sample installed within the sphere for another. For example, when measuring diffuse reflectance (Figure 2, bottom), a first measurement might be made with the standard reference sample mounted inside the sphere. Next, this sample would be removed and replaced by the sample of interest, and a second measurement would be acquired. Both data sets would then be used to calculate the corrected absolute diffuse reflectance spectrum of the sample. 

This procedure would result in a distorted absolute sample spectrum. Since the sample of interest and the standard reference have different absorption and scattering properties, exchanging them alters the reflectivity of the integrating sphere over the samples' surface areas. Due to the average reflectivity of the integrating sphere being different for the two measurements, they are not perfectly compatible.

Solution Option: Install Sample and Reference Together
One experimental technique that avoids sample substitution errors acquires measurement data when both sample and reference are installed inside the integrating sphere at the same time. This approach requires an integrating sphere large enough to accomodate the two, as additional ports. 

The light source is located external to the integrating sphere, and measurements of the sample and standard reference are acquired sequentially. The specular reflection from the sample, or the transmitted beam, is often routed out of the sphere, so that only the diffuse light is detected. Since the inner surface of the sphere is identical for both measurements, sample substitution errors are not a concern.

Alternate Solution Option: Make Measurements from Sample and Reference Ports
If it is not possible to install both sample and standard references in the integrating sphere at the same time, it is necessary to exchange the installed sample. If this must be done, sample substitution errors can be removed by following the procedure detailed in [1].

This procedure requires a total of four measurements. When the standard sample is installed, measurements are made from two different ports. One has a field of view that includes the sample and the other does not. The sample of interest is then subsituted in and the measurements are repeated. Performing the calculations described in [1] using these measurements removes the sample substitution errors.

References
[1] Luka Vidovic and Boris Majaron, "Elimination of single-beam substitution error in diffuse reflectance measurements using an integrating sphere," J. Biomed.Opt. 19, 027006 (2014).

Date of Last Edit: Dec. 4, 2019