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Protocol for whole-mount immunofluorescence staining of ECM gel-embedded innervated pancreatic organoids.
Besikcioglu, Hüseyin Erdinç; Yurteri, Ümmügülsüm; Ye, Linhan; Zhang, Fang; Moretti, Alessandra; Gürcinar, Ibrahim Halil; Dogruöz, Alper; Karakas, Didem; Friess, Helmut; Ceyhan, Guralp Onur; Istvanffy, Rouzanna; Ekin Demir, Ihsan.
Affiliation
  • Besikcioglu HE; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany; Department of Histology and Embryology, Faculty of Medicine, Hitit University, Çorum, Turkey.
  • Yurteri Ü; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Ye L; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Zhang F; Department of Internal Medicine I, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Moretti A; Department of Internal Medicine I, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Gürcinar IH; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Dogruöz A; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Karakas D; Department of Medical Biotechnology, Graduate School of Health Sciences, Acibadem University, Istanbul, Turkey.
  • Friess H; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Ceyhan GO; Department of General Surgery, HPB-Unit, School of Medicine, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey.
  • Istvanffy R; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
  • Ekin Demir I; Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany; Department of General Surgery, HPB-Unit, School of Medicine, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey; German Cancer Consortium (DKTK), Partner Site Munich, Munich,
STAR Protoc ; 5(2): 103132, 2024 Jun 21.
Article in En | MEDLINE | ID: mdl-38875112
ABSTRACT
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.
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Full text: 1 Database: MEDLINE Main subject: Pancreas / Organoids / Fluorescent Antibody Technique / Extracellular Matrix Limits: Animals / Humans Language: En Year: 2024 Type: Article
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Full text: 1 Database: MEDLINE Main subject: Pancreas / Organoids / Fluorescent Antibody Technique / Extracellular Matrix Limits: Animals / Humans Language: En Year: 2024 Type: Article