Light sheet fluorescence microscopy of cleared human eyes

Area of Science:

• Ophthalmology
• Microscopy
• Histology

Background:

• Traditional eye histology methods have limitations in visualizing complex 3D structures.
• There is a need for advanced imaging techniques to study entire human eyes in 3D.

Purpose of the Study:

• To establish a procedure for light sheet fluorescence microscopy (LSFM) of cleared and immunolabeled human eyes.
• To enable 3D navigation and detailed visualization of ocular microanatomy.

Main Methods:

• Human eyes underwent iDISCO+-based clearing (ClearEye) and immunolabeling.
• Light sheet fluorescence microscopy (LSFM) was used to image the entire cleared eyes.
• Post-processing of LSFM data allowed for 3D reconstruction and analysis.

Main Results:

• The method enabled high-resolution imaging of retinal capillaries and complex structures like anterior ciliary vessels and Schlemm's canal.
• Three-dimensional (3D) navigation and customized display, including en face fundus viewing, were achieved.
• The technique was demonstrated successfully on four human eyes.

Conclusions:

• LSFM combined with ClearEye clearing and immunolabeling is a powerful tool for 3D histology of large human ocular samples.
• This method overcomes limitations of traditional histology and facilitates research in anatomopathology and ophthalmology.