Journal article 130 views 22 downloads
Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
M. Carla Carisì,
Owain Howell 
,
Alwena Morgan ,
Jeffrey Davies
,
Alwena Morgan ,
Jeffrey Davies
STAR Protocols, Volume: 3, Issue: 4, Start page: 101896
Swansea University Authors:
Owain Howell , Alwena Morgan , Jeffrey Davies
DOI (Published version): 10.1016/j.xpro.2022.101896
Abstract
We describe a modified BaseScope™ Assay protocol (ACDBio) for RNA in situ hybridization on fixed-frozen human brain tissue. The original protocol caused tissue detachment due to harsh tissue pre-treatment. We therefore optimized it to improve tissue stability while providing high stain quality in fr...
| Published in: | STAR Protocols |
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| ISSN: | 2666-1667 |
| Published: |
Elsevier BV
2022
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| Online Access: |
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa65354 |
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| Abstract: |
We describe a modified BaseScope™ Assay protocol (ACDBio) for RNA in situ hybridization on fixed-frozen human brain tissue. The original protocol caused tissue detachment due to harsh tissue pre-treatment. We therefore optimized it to improve tissue stability while providing high stain quality in fragile post-mortem tissue from aged donors with advanced neurodegeneration. The main changes include two additional fixation steps and modifications to the pre-treatment protocol. We also describe tissue imaging and stain quantification using the open-source QuPath software. |
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| Item Description: |
Protocol |
| College: |
Faculty of Medicine, Health and Life Sciences |
| Issue: |
4 |
| Start Page: |
101896 |