Analyzing live cellularity in the human trabecular meshwork.

ABSTRACT:

Purpose

To directly visualize the live cellularity of the intact human trabecular meshwork (TM) and quantitatively analyze tissue viability in situ.

Methods

Human donor corneoscleral rims were sectioned immediately before intravital dye incubation to label nuclei (Hoechst 33342 & propidium iodide [PI]); cytosol (CellTracker Red CMTPX, calcein AM); and membranes (octadecyl rhodamine B chloride [R18]), followed by 2-photon microscopy. Viability was assessed by counting cells in tissue colabeled with PI and Calcein AM. Some tissues were exposed to Triton X-100 to establish dead tissue controls. Fresh postmortem eyes (within 48 hours of death) represented viable tissue controls. Tissues with live cellularity exceeding 50% were considered viable.

Results

Hoechst nuclear la

SUBMITTER: Gonzalez JM 

PROVIDER: S-EPMC3565993 | biostudies-literature | 2013 Feb

REPOSITORIES: biostudies-literature