Islet Isolation and Cell Sorting

Islet isolation and cell sorting using FACS were performed as described in previous work^{11 (link),62 (link)} using a BD FACSAria II or Moflo Astrios (Beckman Coutler) system. FACSDiva v 8.0.1 (BD Biosciences) software was used for sorting on a FACSAria2, Summit v 6.2 (Beckman Coulter) for sorting on a Moflo Astrios. Kaluza Analysis v 2.0 (Beckman Coulter) software was used for subsequent analysis. Islets were frozen in RLT buffer (Qiagen) with β-Mercaptoethanol and stored at −80 °C before being processed for RNA extraction. RNA extraction from islets was prepared using the Qiagen RNeasy Micro Kit.

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Perez-Frances M., van Gurp L., Abate M.V., Cigliola V., Furuyama K., Bru-Tari E., Oropeza D., Carreaux T., Fujitani Y., Thorel F, & Herrera P.L. (2021). Pancreatic Ppy-expressing γ-cells display mixed phenotypic traits and the adaptive plasticity to engage insulin production. Nature Communications, 12, 4458.

Publication 2021

Moflo Astrios FACSDiva v 8.0.1 Summit v 6.2 RLT buffer RNeasy Micro Kit

Buffer Frozen Isolation Mercaptoethanol

Corresponding Organization :

Other organizations : University of Geneva, Gunma University

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Variable analysis

independent variables

Islet isolation and cell sorting using FACS

dependent variables

RNA extraction from islets

control variables

Islets were frozen in RLT buffer (Qiagen) with β-Mercaptoethanol and stored at −80 °C before being processed for RNA extraction
RNA extraction from islets was prepared using the Qiagen RNeasy Micro Kit

positive controls

Not specified

negative controls

Not specified

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