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High-resolution in-depth imaging of optically cleared thick samples using an adaptive SPIM

Masson, Aurore and Escande, Paul and Frongia, Céline and Clouvel, Grégory and Ducommun, Bernard and Lorenzo, Corinne High-resolution in-depth imaging of optically cleared thick samples using an adaptive SPIM. (2015) Scientific Reports, 5 (1). 1-14. ISSN 2045-2322

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Official URL: http://dx.doi.org/10.1038/srep16898

Abstract

Today, Light Sheet Fluorescence Microscopy (LSFM) makes it possible to image fluorescent samples through depths of several hundreds of microns. However, LSFM also suffers from scattering, absorption and optical aberrations. Spatial variations in the refractive index inside the samples cause major changes to the light path resulting in loss of signal and contrast in the deepest regions, thus impairing in-depth imaging capability. These effects are particularly marked when inhomogeneous, complex biological samples are under study. Recently, chemical treatments have been developed to render a sample transparent by homogenizing its refractive index (RI), consequently enabling a reduction of scattering phenomena and a simplification of optical aberration patterns. One drawback of these methods is that the resulting RI of cleared samples does not match the working RI medium generally used for LSFM lenses. This RI mismatch leads to the presence of low-order aberrations and therefore to a significant degradation of image quality. In this paper, we introduce an original optical-chemical combined method based on an adaptive SPIM and a water-based clearing protocol enabling compensation for aberrations arising from RI mismatches induced by optical clearing methods and acquisition of high-resolution in-depth images of optically cleared complex thick samples such as Multi-Cellular Tumour Spheroids.

Item Type:Article
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Centre National de la Recherche Scientifique - CNRS (FRANCE)
Université de Toulouse > Institut National des Sciences Appliquées de Toulouse - INSA (FRANCE)
Université de Toulouse > Institut Supérieur de l'Aéronautique et de l'Espace - ISAE-SUPAERO (FRANCE)
Other partners > Centre Hospitalier Universitaire de Toulouse - CHU Toulouse (FRANCE)
Other partners > Imagine Optic (FRANCE)
Laboratory name:
Funders:
PRES of Toulouse University and Midi-Pyrenees region
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Deposited By: Paul Escande
Deposited On:25 Apr 2017 13:21

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