Immunostaining of intact C. elegans using polyacrylamide embedding
Summary: A major barrier to immunostaining Caenorhabditis elegans is the permeabilization of the worm's cuticle without distorting or damaging its body. We present here a gel-based immobilization protocol for fixed worms coupled with chemical and enzymatic permeabilization. The permeabilization...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672200836X |
| _version_ | 1811167230708154368 |
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| author | Abbas Ghaddar Wenfan Ke Eyleen J. O’Rourke |
| author_facet | Abbas Ghaddar Wenfan Ke Eyleen J. O’Rourke |
| author_sort | Abbas Ghaddar |
| collection | DOAJ |
| description | Summary: A major barrier to immunostaining Caenorhabditis elegans is the permeabilization of the worm's cuticle without distorting or damaging its body. We present here a gel-based immobilization protocol for fixed worms coupled with chemical and enzymatic permeabilization. The permeabilization is followed by antibody staining and fluorescent imaging. This protocol can be modified for different fixatives, permeabilizing reagents, or molecular readouts. Unlike previous immunostaining approaches, such as freeze cracking or dissection, this protocol enables immunostaining across the whole body of a well-preserved C. elegans. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-10T16:06:56Z |
| format | Article |
| id | doaj.art-47d3610a0b0c469d9ffc617959ce4606 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-10T16:06:56Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-47d3610a0b0c469d9ffc617959ce46062023-02-10T04:23:22ZengElsevierSTAR Protocols2666-16672023-03-0141101956Immunostaining of intact C. elegans using polyacrylamide embeddingAbbas Ghaddar0Wenfan Ke1Eyleen J. O’Rourke2University of Virginia, Department of Biology, Charlottesville, VA 22903, USA; Corresponding authorUniversity of Virginia, Department of Biology, Charlottesville, VA 22903, USAUniversity of Virginia, Department of Biology, Charlottesville, VA 22903, USA; University of Virginia School of Medicine, Department of Cell Biology, Charlottesville, VA 22903, USA; Robert M. Berne Cardiovascular Research Center, School of Medicine, University of Virginia, Charlottesville, VA 22903, USA; Corresponding authorSummary: A major barrier to immunostaining Caenorhabditis elegans is the permeabilization of the worm's cuticle without distorting or damaging its body. We present here a gel-based immobilization protocol for fixed worms coupled with chemical and enzymatic permeabilization. The permeabilization is followed by antibody staining and fluorescent imaging. This protocol can be modified for different fixatives, permeabilizing reagents, or molecular readouts. Unlike previous immunostaining approaches, such as freeze cracking or dissection, this protocol enables immunostaining across the whole body of a well-preserved C. elegans. : 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/S266616672200836XCell BiologyDevelopmental biologyMicroscopyModel OrganismsMolecular BiologyAntibody |
| spellingShingle | Abbas Ghaddar Wenfan Ke Eyleen J. O’Rourke Immunostaining of intact C. elegans using polyacrylamide embedding STAR Protocols Cell Biology Developmental biology Microscopy Model Organisms Molecular Biology Antibody |
| title | Immunostaining of intact C. elegans using polyacrylamide embedding |
| title_full | Immunostaining of intact C. elegans using polyacrylamide embedding |
| title_fullStr | Immunostaining of intact C. elegans using polyacrylamide embedding |
| title_full_unstemmed | Immunostaining of intact C. elegans using polyacrylamide embedding |
| title_short | Immunostaining of intact C. elegans using polyacrylamide embedding |
| title_sort | immunostaining of intact c elegans using polyacrylamide embedding |
| topic | Cell Biology Developmental biology Microscopy Model Organisms Molecular Biology Antibody |
| url | http://www.sciencedirect.com/science/article/pii/S266616672200836X |
| work_keys_str_mv | AT abbasghaddar immunostainingofintactcelegansusingpolyacrylamideembedding AT wenfanke immunostainingofintactcelegansusingpolyacrylamideembedding AT eyleenjorourke immunostainingofintactcelegansusingpolyacrylamideembedding |