June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Assessing biomechanical properties of ex vivo murine retinas using Brillouin microscopy
Author Affiliations & Notes
  • Yogeshwari Sanjayrao Ambekar
    Biomedical Engineering, University of Houston, Houston, Texas, United States
  • Manmohan Singh
    Biomedical Engineering, University of Houston, Houston, Texas, United States
  • Elda M. Rueda
    Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas, United States
  • Benjamin M. Hall
    Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas, United States
  • Jitao Zhang
    Fischell Department of Bioengineering, University of Maryland at College Park, College Park, Maryland, United States
  • Giuliano Scarcelli
    Fischell Department of Bioengineering, University of Maryland at College Park, College Park, Maryland, United States
  • Ross A. Poché
    Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas, United States
  • Kirill Larin
    Biomedical Engineering, University of Houston, Houston, Texas, United States
    Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas, United States
  • Footnotes
    Commercial Relationships   Yogeshwari Sanjayrao Ambekar, None; Manmohan Singh, None; Elda Rueda, None; Benjamin Hall, None; Jitao Zhang, None; Giuliano Scarcelli, None; Ross Poché, None; Kirill Larin, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 1918. doi:
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      Yogeshwari Sanjayrao Ambekar, Manmohan Singh, Elda M. Rueda, Benjamin M. Hall, Jitao Zhang, Giuliano Scarcelli, Ross A. Poché, Kirill Larin; Assessing biomechanical properties of ex vivo murine retinas using Brillouin microscopy. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1918.

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

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Abstract

Purpose : The retina is a crucial part of the eye and plays an important role in vision. Assessing the biomechanical properties of retina could provide critical information for disease detection and guiding precision therapeutic interventions. Previous work assessing retinal elasticity has been limited to global assessments and destructive methods, but in this work we demonstrate Brillouin microscopy to noninvasively assess the layer by layer distribution of retinal stiffness. Furthermore, we were able to measure the changes in retinal stiffness caused by fixation and the effects of n-methyl-d-aspartate (NMDA) induced damage.

Methods : Dissected fresh (n = 3) and paraformaldehyde-fixed (n = 2) adult C57/BL6J mice retinas were used in this experiment. The retinas were mounted flat on microscope slides with the vitreous layer on the top and the photoreceptor layer on the bottom. Optical coherence tomography (OCT) images were acquired to visualize the retina structure and ensure there was no damage. Next, Brillouin microscopy was used to obtain the layer by layer distribution of biomechanical properties of the retinas. As a proof-of-concept study, mice were intravitreally injected with 100 mM NMDA and imaged using OCT and Brillouin microscopy.

Results : We observed that the average Brillouin modulus of all fresh retinas over entire depth was 2.48±0.06 GPa whereas the average Brillouin modulus of all fixed retinas was 2.73±0.09 GPa. Figure 1 (a) shows layer by layer distribution of stiffness within the retina using Brillouin microscopy. Retinal ganglion cell (RGC) layer of NMDA-damaged retina had a greater Brillouin modulus compared to the control retina sample.

Conclusions : We showed that Brillouin microscopy can noninvasively assess the stiffness over the entire depth of ex vivo mouse retinas. We observed that the retina is a heterogeneous tissue and has different biomechanical properties for its different layers. Fixed retinas were stiffer compared to fresh retinas. Results showed the biomechanical properties are highly affected by the cellular density and PFA-fixing. Increased Brillouin modulus in the RGC layer of NMDA-induced retina may indicate cell death in that layer.

This is a 2021 ARVO Annual Meeting abstract.

 

Figure 1 (a) Layer by layer distribution of stiffness in terms of Brillouin modulus. (b) Average Brillouin modulus of PFA-fixed (n=2) and fresh (n=3) mouse retina (c) Average Brillouin modulus of RGC layer of control and NMDA-induced damage retinas

Figure 1 (a) Layer by layer distribution of stiffness in terms of Brillouin modulus. (b) Average Brillouin modulus of PFA-fixed (n=2) and fresh (n=3) mouse retina (c) Average Brillouin modulus of RGC layer of control and NMDA-induced damage retinas

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