June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Toward longitudinal evaluations of biological processes in transgenic mice models using an integrated cellular-resolution optical coherence microscopy and fluorescence microscopy.
Author Affiliations & Notes
  • Reddikumar Maddipatla
    school of Optometry, Indiana University Bloomington, Bloomington, Indiana, United States
  • Chia-Yang Liu
    school of Optometry, Indiana University Bloomington, Bloomington, Indiana, United States
  • Patrice Tankam
    school of Optometry, Indiana University Bloomington, Bloomington, Indiana, United States
  • Footnotes
    Commercial Relationships   Reddikumar Maddipatla None; Chia-Yang Liu None; Patrice Tankam None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 107 – A0205. doi:
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      Reddikumar Maddipatla, Chia-Yang Liu, Patrice Tankam; Toward longitudinal evaluations of biological processes in transgenic mice models using an integrated cellular-resolution optical coherence microscopy and fluorescence microscopy.. Invest. Ophthalmol. Vis. Sci. 2022;63(7):107 – A0205.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Nowadays, scientists are equipped with robust tools to induce genetic mutations in animal models to advance the understanding of disease mechanisms. The purpose of this work is to enable the evaluation of the outcomes of these genetic modifications and follow up these processes over time on a single specimen using advanced optical imaging techniques.

Methods : An integrated optical coherence microscopy (OCM) and dual-channel fluorescence microscopy (DCFM) system was developed to simultaneously co-register the reflectance and fluorescence signals from the cornea of transgenic mouse models. OCM was equipped with a broadband source with a central wavelength at 850 nm and FWHM of 165 nm. DCFM was designed with two excitation peaks at 473 nm for Tdtomato and 561 nm for green fluorescent proteins (GFP). A conditional knockout mouse that expresses both tdTomato and GFP in the corneal stroma was engineered to evaluate the system performances. Reflectance and fluorescence signals were registered simultaneously at a given plane of focus in the corneal stroma. However, unlike OCM, DCFM does not possess an intrinsic depth-sectioning capability. Therefore, subsequent acquisitions with DCFM alone were performed by z-scanning the sample with a step size of 2 µm to evaluate the axial resolution of DCFM.

Results : The system achieved the simultaneous co-registration of reflectance and fluorescence signals with a lateral resolution of 2 µm and speed of 250kHz line rate. OCM achieved an axial resolution of ~2 µm in the cornea.

Conclusions : The custom multimodal system was evaluated on corneal imaging of a transgenic mouse. Future work will focus on enhancing the contrast detection and depth-sectioning of DCFM.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

Fig.1 Images of a transgenic mouse cornea. A-C Cross-sectional images of OCM, green, and red DCFM. E-G En face images at the plane of focus (blue line in A-D). D, H Composite images of OCM, green, and red DCFM.

Fig.1 Images of a transgenic mouse cornea. A-C Cross-sectional images of OCM, green, and red DCFM. E-G En face images at the plane of focus (blue line in A-D). D, H Composite images of OCM, green, and red DCFM.

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