Cellular assessment of the cornea of transgenic mice models using multi-modal optical coherence microscopy and dual-channel fluorescence microscopy

Reddikumar Maddipatla; Xiao Liu; Chia-Yang Liu; Patrice Tankam

doi:10.1117/12.2610212

2 March 2022 Cellular assessment of the cornea of transgenic mice models using multi-modal optical coherence microscopy and dual-channel fluorescence microscopy

Reddikumar Maddipatla, Xiao Liu, Chia-Yang Liu, Patrice Tankam

Author Affiliations +

Proceedings Volume 11952, Multimodal Biomedical Imaging XVII; 1195209 (2022) https://doi.org/10.1117/12.2610212
Event: SPIE BiOS, 2022, San Francisco, California, United States

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Abstract

Scientists today have powerful tools to alter a wide range of genes in animal models with ease to understand biological processes in development and disease conditions. However, there is a lack of imaging technology to assess the effects of these genetic modifications in live mammals. Without these in vivo imaging capabilities, scientists cannot fully understand how the individual organism’s response to overall process. In this work, we have developed a multi-modal optical coherence microscopy (OCM) and dual-channel fluorescence microscopy (DC-FM) to enable the evaluation of biological processes in transgenic mice models. The combined system achieved a simultaneous recording of reflectance and fluorescence signals of deep tissue layers at the speed of 250 kHz and a lateral resolution of ~ 2 μm over a field of view of 1.3 x 1.3 mm² . OCM and DC-FM also achieved an axial resolution of 2.8 μm and 23 μm, respectively. To evaluate the performances of the system in imaging the cornea of transgenic mice, a conditional dual-reporter mouse strain, KerartTA/tet-O-Cre/RosamTmG triple transgenic mouse strain, which express membrane-bound tomato red (mT), was harnessed. Upon exposure to doxycycline (Dox) in KerartTA/tet-O-Cre/RosamTmG mice, it would then express a membrane-bound green fluorescent protein (mG) in the corneal stromal cells. While OCM evaluated the change in thickness and structures of the cornea, DC-FM provided green and red proteins expression in the cornea. This system will enable longitudinal in vivo studies on transgenic mice models to advance the understanding of developmental and disease mechanisms.

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Reddikumar Maddipatla, Xiao Liu, Chia-Yang Liu, and Patrice Tankam "Cellular assessment of the cornea of transgenic mice models using multi-modal optical coherence microscopy and dual-channel fluorescence microscopy", Proc. SPIE 11952, Multimodal Biomedical Imaging XVII, 1195209 (2 March 2022); https://doi.org/10.1117/12.2610212

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