Current limitations in super-resolution fluorescence microscopy for biological specimens: How deep can we go from the cover glass?

Yasushi Okada

doi:10.1117/12.2276015

18 April 2017 Current limitations in super-resolution fluorescence microscopy for biological specimens: How deep can we go from the cover glass?

Yasushi Okada

Author Affiliations +

Proceedings Volume 10251, Biomedical Imaging and Sensing Conference; 102510I (2017) https://doi.org/10.1117/12.2276015
Event: SPIE Technologies and Applications of Structured Light, 2017, Yokohama, Japan

Abstract

Diffraction limit of resolution has been one of the biggest limitations in the optical microscopy. Super-resolution fluorescence microscopy has enabled us to break this limit. However, for the observations of real biological specimens, especially for the imaging of tissues or whole body, the target structures of interest are often embedded deep inside the specimen. Here, we would present our results to extend the target of the super-resolution microscopy deeper into the cells. Confocal microscope optics work effectively to minimize the effect by the aberrations by the cellular components, but at the expense of the signal intensities. Spherical aberrations by the refractive index mismatch between the cellular environment and the immersion liquid can be much larger, but can be reduced by adjusting the correction collar at the objective lens.

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Yasushi Okada "Current limitations in super-resolution fluorescence microscopy for biological specimens: How deep can we go from the cover glass?", Proc. SPIE 10251, Biomedical Imaging and Sensing Conference, 102510I (18 April 2017); https://doi.org/10.1117/12.2276015

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