Protocol for whole-mount immunofluorescence staining of ECM gel-embedded innervated pancreatic organoids

Besikcioglu, Huseyin; Yurteri, Ummugulsum; Ye, Linhan; Zhang, Fang; Moretti, Alessandra; Guercinar, Ibrahim; Dogruoez, Alper; KARAKAŞ ZEYBEK, DİDEM; Friess, Helmut; CEYHAN, GÜRALP; Istvanffy, Rouzanna; Demir, Ihsan

doi:10.1016/j.xpro.2024.103132

Protocol for whole-mount immunofluorescence staining of ECM gel-embedded innervated pancreatic organoids

Besikcioglu H. E., Yurteri U., Ye L., Zhang F., Moretti A., Guercinar I. H., ...Daha Fazla

STAR PROTOCOLS, cilt.5, sa.2, 2024 (ESCI, Scopus)

Yayın Türü: Makale / Tam Makale
Cilt numarası: 5 Sayı: 2
Basım Tarihi: 2024
Doi Numarası: 10.1016/j.xpro.2024.103132
Dergi Adı: STAR PROTOCOLS
Derginin Tarandığı İndeksler: Emerging Sources Citation Index (ESCI), Scopus
Acıbadem Mehmet Ali Aydınlar Üniversitesi Adresli: Evet

Özet

The mandatory usage of extracellular matrix (ECM) gels in 3D cultures limits antibody penetration and increases background, while the removal of ECM gel causes disruption of morphology and sample loss. These factors pose challenges to effective immune labeling -based staining. Here, we present a protocol for whole -mount immunofluorescence staining of gel -embedded pancreatic organoids. We describe steps for sample fixation, blocking, and antibody incubation. We detail procedures for washing antibodies and mounting.