Retinal Microvasculature by 3D Confocal Microscopy

Immunofluorescence imaged with a Zeiss LSM510 Meta confocal microscope. C57/BL6 mouse eye cups were incubated with Mouse monoclonal anti-Glutamine Synthetase (Chemicon, MAB302) to detect Muller Cells and visualized using donkey anti-mouse IgG Alexafluor 488 (Invitrogen, A21202). Vascular endothelial cells were detected using Isolectin B4, Griffonia simplicifolia, conjugated with Alexafluor 568 (Invitrogen, I21412). Nuclei were probed using Hoechst 33258. Following immunolabeling, tissue was embedded in Epon-araldite expoxy, faced with a Leica RM 2165 microtome, and floated on a 40x/NA1.4 oil immersion objective. Rotating 3D retinas are oriented so that the vitreal surface is down.

Category:

Science & Technology

Tags:

• Retina
• Microvsculature
• 3D Reconstruction
• Isolectin
• Glutamine Synthetase
• Muller Cells
• Vascular Endothelial Cells
• Bazan Laboratory