Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue

Lydia Farack; Shalev Itzkovitz

doi:https://doi.org/10.1016/j.xpro.2019.100007

Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue

Lydia Farack, Shalev Itzkovitz

Department of Molecular Cell Biology

Weizmann Institute of Science

Research output: Contribution to journal › Article › peer-review

Abstract

We describe an optimized smFISH protocol for the intact pancreas. The protocol is adapted from Lyubimova et al. (2013), a generic tissue smFISH protocol that works for most tissues but not the pancreas. The main changes implemented include increasing the period of mRNA denaturation from 5 min to at least 3 h and increasing formamide concentrations from 10% to 30%. These modifications yield sensitive single mRNA visualization that is comparable to those achieved in other tissues using the standard protocol. For complete details on the use and execution of this protocol, please refer to Farack et al., 2018, Farack et al., 2019.

Original language	English
Article number	100007
Number of pages	19
Journal	STAR Protocols
Volume	1
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2019.100007
State	Published - 19 Jun 2020

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si_Star-Pro_Protocol-for-single-molecule_PV_2020Final published version, 3.61 MBLicense: CC BY-NC-ND

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title = "Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue",

abstract = "We describe an optimized smFISH protocol for the intact pancreas. The protocol is adapted from Lyubimova et al. (2013), a generic tissue smFISH protocol that works for most tissues but not the pancreas. The main changes implemented include increasing the period of mRNA denaturation from 5 min to at least 3 h and increasing formamide concentrations from 10% to 30%. These modifications yield sensitive single mRNA visualization that is comparable to those achieved in other tissues using the standard protocol. For complete details on the use and execution of this protocol, please refer to Farack et al., 2018, Farack et al., 2019.",

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Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue

Abstract

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