Home Microscopy & Imaging Full-Field Optical Coherence Microscopy for Histology-Like Analysis of Stromal Features in Corneal Grafts
Microscopy & Imaging JoVE (Open Access) Citable · DOI

Full-Field Optical Coherence Microscopy for Histology-Like Analysis of Stromal Features in Corneal Grafts

DOI: 10.3791/57104-v
What you'll learn
  • Prepare corneal donor tissue samples for full-field optical coherence microscopy imaging
  • Operate FF-OCM system to acquire high-resolution stromal images without histological processing
  • Perform qualitative and quantitative analysis of corneal stroma morphology and disease features
  • Apply imaging results to improve donor tissue screening for keratoplasty
Protocol

We describe use of full-field optical coherence microscopy as a method for high quality assessment of corneal donor stroma. This protocol can be used to identify features indicative of health or disease, and is aimed at improving the screening and selection of donor tissues, and hence the results of keratoplasty.

Difficulty
advanced
Total time
~1–2 hours per corneal graft sample (including setup, imaging, and analysis)

Steps

1
Understand full-field OCM principles and clinical applications

Review optical coherence microscopy theory, advantages for non-destructive corneal assessment, and relevance to donor tissue evaluation and keratoplasty outcomes.

▶ 00:04
2
Prepare corneal donor tissue for imaging

Mount and orient the donor corneal graft sample on the FF-OCM stage according to protocol specifications for optimal stromal visualization.

▶ 00:47
3
Initialize system and acquire stromal images

Calibrate the FF-OCM instrument, set imaging parameters, and acquire high-resolution cross-sectional images of the corneal stroma.

▶ 01:31
4
Analyze stromal morphology qualitatively and quantitatively

Examine acquired images for structural features (collagen organization, edema, scarring) and perform quantitative measurements to assess tissue health and disease indicators.

▶ 03:48
5
Interpret results and assess donor tissue quality

Synthesize qualitative and quantitative findings to characterize donor stroma status and inform tissue suitability for transplantation.

▶ 06:08
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