Trans-scleral imaging of the human trabecular meshwork by two-photon microscopy

[摘要] Purpose: To image the native (unfixed)human trabecular meshwork (TM) through the overlying sclera using anon-invasive, non-destructive technique. Methods: Two-photon microscopic (2PM)methods, including two-photon autofluorescence (2PAF) and secondharmonic generation (SHG), were used to image through the sclera of ahuman cadaver eye into the TM region. Multiple imageswere analyzed along the tissue axis (z-axis) to generate athree-dimensional (3D) model of the region. The tissue was subsequentlyfixed, paraffin embedded, and histological sections were photographedfor comparison to the 2PM images. Results: 3D analysis of multiple 2PM SHGimages revealed an open region deep within the TM consistent with thelocation of Schlemm’s canal (SC). Images of the scleral spur andsurrounding tissues were also obtained. The SC, TM, scleral spur, andsurrounding tissue images obtained with 2PM matched with histologicallystained sections of the same tissue. Conclusions: 2PM imaging of the outflowsystem of the human eye documented collagenous structures solely frominherent optical properties. 2PM successfully imaged through the sclerainto the SC/TM without the need for fixation, embedding, orhistological processing. This work reveals that 2PM imaging haspotential as a new metric for evaluating the aqueous outflow region ofthe human eye and is worthy of further exploration.

[学科分类] 生物化学/生物物理