Protocol to quantify enzymatic effects on vitreous liquefaction in porcine eyes using a transwell-plate system

doi:10.1016/j.xpro.2022.101754

STAR Protocols

Volume 3, Issue 4, 16 December 2022, 101754

https://doi.org/10.1016/j.xpro.2022.101754 Get rights and content

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open access

Highlights

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Transwell-plate-based protocol for quantifying vitreous liquefaction ex vivo
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Perform vitreous separation through a process of induced syneresis
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Perform multiple experiments from a porcine eye with minimal sample loss
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Experimental design allows end-product analysis to account for sample loss

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Summary

This protocol describes an ex vivo model to quantify enzymatic effects on vitreous liquefaction using porcine eyes in a transwell-plate system via induced syneresis. It provides a standardized dissection process and performs critical steps for gel-liquid separation with high precision, minimal tissue loss, and scalability. The protocol can be applied to other studies investigating vitreous liquefaction or gelatinous tissue analysis and can also serve to study vitreous liquefaction in vivo as it may occur during aging or disease progression.

Graphical abstract

Subject areas

Health sciences

Molecular biology

Protein biochemistry

Data and code availability

All data generated in this paper will be shared by the lead contact by request.

Any additional information required to reanalyze the data reported in this work is available from the lead contact upon request.

Cited by (0)

⁴: Technical contact: [email protected]

⁵: Lead contact