Corneal Cell Isolation and Analysis

A single-cell suspension of corneal cells was prepared by digesting the individual cornea with Liberase TL (2.5 mg/ml) (Sigma-Aldrich) as previously reported (Gaddipati et al., 2015 (link)). Cells were stained with fluorochrome-conjugated monoclonal antibodies against CD45 (103116, Biolegend), HIF-1α (IC1935G, R&D system) and their isotype controls (400623, 400132, Biolegend). The stained cells were analyzed using the LSRII flow cytometer (BD Biosciences, San Jose, CA) and FlowJo software.

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Zhu S., Shan H., Li J., Pan L., Wang S., Zhu J., Guo H., Mi F., Wu X., Yin J, & Pang K. (2022). Therapeutic potential of topical administration of acriflavine against hypoxia-inducible factors for corneal fibrosis. Frontiers in Pharmacology, 13, 996635.

Publication 2022

Liberase TL IC1935G LSRII flow cytometer

Cells Cornea Fluorochrome Isotype Liberase Monoclonal antibodies

Corresponding Organization : Smith-Kettlewell Eye Research Institute

Other organizations : Shanxi Eye Hospital

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Variable analysis

independent variables

Digestion of corneal cells with Liberase TL (2.5 mg/ml)

dependent variables

Expression of CD45
Expression of HIF-1α

control variables

Isotype controls for CD45 and HIF-1α

controls

Positive control: Not specified
Negative control: Isotype controls for CD45 and HIF-1α

Annotations

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