Retinal diseases such as diabetic retinopathy and glaucoma are leading cause of blindness in the world. Assessing biomechanical properties of retina is very crucial since it is constantly under stress due to the vitreous humor and eye movements. Characterizing biomechanical properties of retina noninvasively is a challenge due to its location inside the eye-globe, fragility, and thin geometry. Brillouin microscopy is a noninvasive, all optical imaging technique to qualitatively map the biomechanical properties of tissues. In this work, we mapped the layer by layer distribution of biomechanical properties of retinas using Brillouin microscopy. We found that the nuclear layer was stiffer compared to other layers. Furthermore, we observed fixing the retinas with paraformaldehyde increased the retinal stiffness compared to the fresh retinas.
Significance: The retina is critical for vision, and several diseases may alter its biomechanical properties. However, assessing the biomechanical properties of the retina nondestructively is a challenge due to its fragile nature and location within the eye globe. Advancements in Brillouin spectroscopy have provided the means for nondestructive investigations of retina biomechanical properties.
Aim: We assessed the biomechanical properties of mouse retinas using Brillouin microscopy noninvasively and showed the potential of Brillouin microscopy to differentiate the type and layers of retinas based on stiffness.
Approach: We used Brillouin microscopy to quantify stiffness of fresh and paraformaldehyde (PFA)-fixed retinas. As further proof-of-concept, we demonstrated a change in the stiffness of a retina with N-methyl-D-aspartate (NMDA)-induced damage, compared to an undamaged sample.
Results: We found that the retina layers with higher cell body density had higher Brillouin modulus compared to less cell-dense layers. We have also demonstrated that PFA-fixed retina samples were stiffer compared with fresh samples. Further, NMDA-induced neurotoxicity leads to retinal ganglion cell (RGC) death and reactive gliosis, increasing the stiffness of the RGC layer.
Conclusion: Brillouin microscopy can be used to characterize the stiffness distribution of the layers of the retina and can be used to differentiate tissue at different conditions based on biomechanical properties.
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