Protocol for immunofluorescence characterization of drusen in induced pluripotent stem cell-derived retinal pigment epithelium cultures
Drusen, lipoprotein-rich deposits underlying retinal pigment epithelium (RPE), are a pathological hallmark of age-related macular degeneration and related macular dystrophies. Here, we describe a protocol for visualizing drusen deposits beneath induced pluripotent stem cell (iPSC)-RPE (iRPE) monocultures. We describe steps for iRPE cell culture and, subsequently, enzymatic removal of the iRPE monolayer and immunostaining of sub-RPE drusen deposits. We then detail steps for confocal microscopy imaging of immunostained drusen in iRPE culture. For complete details on the use and execution of this protocol, please refer to Dalvi et al.<sup>1</sup>.
Authors
- Sonal Dalvi
- Ruchira Singh (ORCID: https://orcid.org/0000-0002-3821-1538)
Institutions
- University of Rochester Medical Center (US)
- University of Rochester (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-05-02
- DOI
- https://doi.org/10.1016/j.xpro.2025.103791
- Primary Topic
- Retinal Diseases and Treatments
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Research to Prevent Blindness
- National Institutes of Health
- School of Medicine and Dentistry, University of Rochester
- National Eye Institute