Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy
Summary: Due to their small size and transparency, larval zebrafish are a useful model for whole-brain imaging. Here, we present a protocol for the visualization of phosphorylated Rps6, a marker of mechanistic target of rapamycin complex 1 (mTORC1) activity, in the zebrafish brains at 5 days post fe...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007524 |
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| author | Olga Doszyn Tomasz Dulski Justyna Zmorzynska |
| author_facet | Olga Doszyn Tomasz Dulski Justyna Zmorzynska |
| author_sort | Olga Doszyn |
| collection | DOAJ |
| description | Summary: Due to their small size and transparency, larval zebrafish are a useful model for whole-brain imaging. Here, we present a protocol for the visualization of phosphorylated Rps6, a marker of mechanistic target of rapamycin complex 1 (mTORC1) activity, in the zebrafish brains at 5 days post fertilization (dpf), using whole-mount immunofluorescence and light-sheet microscopy. We describe steps for sample preparation, storage, staining, and imaging. This protocol can also be modified for staining with antibodies against other proteins.For complete details on the use and execution of this protocol, please refer to Doszyn et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-d659731902dd4d49aa2ceb217c06729d |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-d659731902dd4d49aa2ceb217c06729d2025-01-19T06:26:41ZengElsevierSTAR Protocols2666-16672025-03-0161103587Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopyOlga Doszyn0Tomasz Dulski1Justyna Zmorzynska2Laboratory of Developmental Neurobiology, International Institute of Molecular Mechanisms and Machines, 02-247 Warsaw, Poland; Laboratory of Molecular and Cellular Neurobiology, International Institute of Molecular and Cell Biology in Warsaw, 02-109 Warsaw, PolandLaboratory of Developmental Neurobiology, International Institute of Molecular Mechanisms and Machines, 02-247 Warsaw, PolandLaboratory of Developmental Neurobiology, International Institute of Molecular Mechanisms and Machines, 02-247 Warsaw, Poland; Laboratory of Molecular and Cellular Neurobiology, International Institute of Molecular and Cell Biology in Warsaw, 02-109 Warsaw, Poland; Corresponding authorSummary: Due to their small size and transparency, larval zebrafish are a useful model for whole-brain imaging. Here, we present a protocol for the visualization of phosphorylated Rps6, a marker of mechanistic target of rapamycin complex 1 (mTORC1) activity, in the zebrafish brains at 5 days post fertilization (dpf), using whole-mount immunofluorescence and light-sheet microscopy. We describe steps for sample preparation, storage, staining, and imaging. This protocol can also be modified for staining with antibodies against other proteins.For complete details on the use and execution of this protocol, please refer to Doszyn et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724007524developmental biologymicroscopymodel organismsneuroscience |
| spellingShingle | Olga Doszyn Tomasz Dulski Justyna Zmorzynska Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy STAR Protocols developmental biology microscopy model organisms neuroscience |
| title | Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy |
| title_full | Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy |
| title_fullStr | Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy |
| title_full_unstemmed | Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy |
| title_short | Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy |
| title_sort | protocol for the visualization of prps6 positive cells in larval zebrafish brains using whole mount immunofluorescence and light sheet microscopy |
| topic | developmental biology microscopy model organisms neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007524 |
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