Home > Publications database > Method for the refractive index of various tissues based on fluorescence microscopy |
Journal Article | FZJ-2023-03083 |
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2023
Optica Publishing Group
Washington, DC
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Please use a persistent id in citations: doi:10.1364/OPTCON.492897 doi:10.34734/FZJ-2023-03083
Abstract: Refractive index is an important optical constant that characterizes the interactionbetween light and specimen. A difference in refractive index between specimen and immersionmedium introduces the imaging aberration and leads to a problem that the direct thicknessmeasurement of a specimen by optical microscopy is not accurate. However, this aberrationcorrection still requires the exact information of the refractive index of specimen and immersionmedium. Herein, we propose an imaging strategy to estimate the refractive index for an unknownspecimen. A simplified diffraction model is generated to obtain the relationship between axialscaling factor and refractive index. Then regular fluorescence microscopy is performed to measurethe actual axial scaling factors of specimens from mouse muscle and tumor xenograft. Referringto our theoretical plot of axial scaling factor versus refractive index, the refractive index of tissuespecimen is determined. For example, we obtain a mean refractive index (n) value of 1.36 fornormal muscle tissues, and 1.41 for tumor xenografts. We demonstrate that this diffractionmodel-based estimation method is an alternative to the current techniques, improving the accuratemeasurement for refractive index of tissue specimen. The simple instrument requirement with aneasy specimen preparation for this estimation method of refractive index may increase the imagequality on tissue specimens with less aberration.
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