Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
Summary: 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 m...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | STAR Protocols |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166719300073 |
| _version_ | 1811292309772304384 |
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| author | Lydia Farack Shalev Itzkovitz |
| author_facet | Lydia Farack Shalev Itzkovitz |
| author_sort | Lydia Farack |
| collection | DOAJ |
| description | Summary: 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, 2019). |
| first_indexed | 2024-04-13T04:43:31Z |
| format | Article |
| id | doaj.art-91b10f0262cc4a969127806566b33815 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T04:43:31Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-91b10f0262cc4a969127806566b338152022-12-22T03:01:56ZengElsevierSTAR Protocols2666-16672020-06-0111100007Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic TissueLydia Farack0Shalev Itzkovitz1Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel; Corresponding authorDepartment of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel; Corresponding authorSummary: 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, 2019).http://www.sciencedirect.com/science/article/pii/S2666166719300073 |
| spellingShingle | Lydia Farack Shalev Itzkovitz Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue STAR Protocols |
| title | Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue |
| title_full | Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue |
| title_fullStr | Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue |
| title_full_unstemmed | Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue |
| title_short | Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue |
| title_sort | protocol for single molecule fluorescence in situ hybridization for intact pancreatic tissue |
| url | http://www.sciencedirect.com/science/article/pii/S2666166719300073 |
| work_keys_str_mv | AT lydiafarack protocolforsinglemoleculefluorescenceinsituhybridizationforintactpancreatictissue AT shalevitzkovitz protocolforsinglemoleculefluorescenceinsituhybridizationforintactpancreatictissue |