Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development
Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of...
Gespeichert in:
| Veröffentlicht in: | Journal of Visualized Experiments 2014-05, Vol.87 (87) |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext bestellen |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | |
|---|---|
| container_issue | 87 |
| container_start_page | |
| container_title | Journal of Visualized Experiments |
| container_volume | 87 |
| creator | Chardès, Claire Mélénec, Pauline Bertrand, Vincent Lenne, Pierre-François |
| description | Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time. |
| doi_str_mv | 10.3791/51342 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_223</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4172305</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1526735904</sourcerecordid><originalsourceid>FETCH-LOGICAL-c486t-4c4491594cfc7ad3ae2576f69ca2d878a09b7c041e06bfcc7694c620defe87e43</originalsourceid><addsrcrecordid>eNpdkV9PwjAUxRujEUS-gA-mLyb6MGzXbt1eTAj-gQTjA5Dw1pTuboxs61wHid_eTZCgT216f-fc23sQ6lMyYCKkjx5l3D1DXRpy4pBALM9P7h10Ze2GEN8lXnCJOi4PmM-J6KLlDOo6LRK8KLHCszQvM8DTNFnXeLYGqPF7qitjtSkBx6bCkwLPTW3wJFdJKzMxHg0wZJCowuJn2EFmyhyK-hpdxCqz0D-cPbR4fZmPxs70420yGk4dzQO_drjmPKReyHWshYqYAtcTfuyHWrlRIAJFwpXQhFMg_irWWvgN2nwjghgCAZz10NPet9yucoh007pSmSyrNFfVlzQqlX8rRbqWidlJToXLiNcYPOwN1v9k4-FUtm-EUsIZ83a0Ye8PzSrzuQVbyzy1GrJMFWC2VlLP9QXzQtLOdbdH2_XZCuKjNyWyTUz-JNZwt6fzH6nfiBrgZg9szA7kxmyrotnnQf0N7eaYIg</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1526735904</pqid></control><display><type>article</type><title>Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development</title><source>Journal of Visualized Experiments : JoVE</source><creator>Chardès, Claire ; Mélénec, Pauline ; Bertrand, Vincent ; Lenne, Pierre-François</creator><creatorcontrib>Chardès, Claire ; Mélénec, Pauline ; Bertrand, Vincent ; Lenne, Pierre-François</creatorcontrib><description>Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time.</description><identifier>ISSN: 1940-087X</identifier><identifier>EISSN: 1940-087X</identifier><identifier>DOI: 10.3791/51342</identifier><identifier>PMID: 24836407</identifier><language>eng</language><publisher>United States: MyJove Corporation</publisher><subject>Animal biology ; Animals ; Caenorhabditis elegans - embryology ; Cellular Biology ; Developmental Biology ; Embryo, Nonmammalian ; Imaging, Three-Dimensional - instrumentation ; Imaging, Three-Dimensional - methods ; Life Sciences ; Microscopy - instrumentation ; Microscopy - methods ; Time-Lapse Imaging - instrumentation ; Time-Lapse Imaging - methods</subject><ispartof>Journal of Visualized Experiments, 2014-05, Vol.87 (87)</ispartof><rights>Copyright © 2014, Journal of Visualized Experiments</rights><rights>Attribution - NonCommercial - NoDerivatives</rights><rights>Copyright © 2014, Journal of Visualized Experiments 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c486t-4c4491594cfc7ad3ae2576f69ca2d878a09b7c041e06bfcc7694c620defe87e43</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttps://www.jove.com/files/email_thumbs/51342.png</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4172305/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4172305/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,3843,27924,27925,53791,53793</link.rule.ids><linktorsrc>$$Uhttp://dx.doi.org/10.3791/51342$$EView_record_in_Journal_of_Visualized_Experiments$$FView_record_in_$$GJournal_of_Visualized_Experiments</linktorsrc><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24836407$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-01104335$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Chardès, Claire</creatorcontrib><creatorcontrib>Mélénec, Pauline</creatorcontrib><creatorcontrib>Bertrand, Vincent</creatorcontrib><creatorcontrib>Lenne, Pierre-François</creatorcontrib><title>Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development</title><title>Journal of Visualized Experiments</title><addtitle>J Vis Exp</addtitle><description>Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time.</description><subject>Animal biology</subject><subject>Animals</subject><subject>Caenorhabditis elegans - embryology</subject><subject>Cellular Biology</subject><subject>Developmental Biology</subject><subject>Embryo, Nonmammalian</subject><subject>Imaging, Three-Dimensional - instrumentation</subject><subject>Imaging, Three-Dimensional - methods</subject><subject>Life Sciences</subject><subject>Microscopy - instrumentation</subject><subject>Microscopy - methods</subject><subject>Time-Lapse Imaging - instrumentation</subject><subject>Time-Lapse Imaging - methods</subject><issn>1940-087X</issn><issn>1940-087X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkV9PwjAUxRujEUS-gA-mLyb6MGzXbt1eTAj-gQTjA5Dw1pTuboxs61wHid_eTZCgT216f-fc23sQ6lMyYCKkjx5l3D1DXRpy4pBALM9P7h10Ze2GEN8lXnCJOi4PmM-J6KLlDOo6LRK8KLHCszQvM8DTNFnXeLYGqPF7qitjtSkBx6bCkwLPTW3wJFdJKzMxHg0wZJCowuJn2EFmyhyK-hpdxCqz0D-cPbR4fZmPxs70420yGk4dzQO_drjmPKReyHWshYqYAtcTfuyHWrlRIAJFwpXQhFMg_irWWvgN2nwjghgCAZz10NPet9yucoh007pSmSyrNFfVlzQqlX8rRbqWidlJToXLiNcYPOwN1v9k4-FUtm-EUsIZ83a0Ye8PzSrzuQVbyzy1GrJMFWC2VlLP9QXzQtLOdbdH2_XZCuKjNyWyTUz-JNZwt6fzH6nfiBrgZg9szA7kxmyrotnnQf0N7eaYIg</recordid><startdate>20140505</startdate><enddate>20140505</enddate><creator>Chardès, Claire</creator><creator>Mélénec, Pauline</creator><creator>Bertrand, Vincent</creator><creator>Lenne, Pierre-François</creator><general>MyJove Corporation</general><general>JoVE</general><scope>ALOKQ</scope><scope>DRUMS</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><scope>5PM</scope></search><sort><creationdate>20140505</creationdate><title>Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development</title><author>Chardès, Claire ; Mélénec, Pauline ; Bertrand, Vincent ; Lenne, Pierre-François</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c486t-4c4491594cfc7ad3ae2576f69ca2d878a09b7c041e06bfcc7694c620defe87e43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animal biology</topic><topic>Animals</topic><topic>Caenorhabditis elegans - embryology</topic><topic>Cellular Biology</topic><topic>Developmental Biology</topic><topic>Embryo, Nonmammalian</topic><topic>Imaging, Three-Dimensional - instrumentation</topic><topic>Imaging, Three-Dimensional - methods</topic><topic>Life Sciences</topic><topic>Microscopy - instrumentation</topic><topic>Microscopy - methods</topic><topic>Time-Lapse Imaging - instrumentation</topic><topic>Time-Lapse Imaging - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chardès, Claire</creatorcontrib><creatorcontrib>Mélénec, Pauline</creatorcontrib><creatorcontrib>Bertrand, Vincent</creatorcontrib><creatorcontrib>Lenne, Pierre-François</creatorcontrib><collection>JoVE Journal: Biology</collection><collection>JoVE Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Visualized Experiments</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Chardès, Claire</au><au>Mélénec, Pauline</au><au>Bertrand, Vincent</au><au>Lenne, Pierre-François</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development</atitle><jtitle>Journal of Visualized Experiments</jtitle><addtitle>J Vis Exp</addtitle><date>2014-05-05</date><risdate>2014</risdate><volume>87</volume><issue>87</issue><issn>1940-087X</issn><eissn>1940-087X</eissn><abstract>Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time.</abstract><cop>United States</cop><pub>MyJove Corporation</pub><pmid>24836407</pmid><doi>10.3791/51342</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext_linktorsrc |
| identifier | ISSN: 1940-087X |
| ispartof | Journal of Visualized Experiments, 2014-05, Vol.87 (87) |
| issn | 1940-087X 1940-087X |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4172305 |
| source | Journal of Visualized Experiments : JoVE |
| subjects | Animal biology Animals Caenorhabditis elegans - embryology Cellular Biology Developmental Biology Embryo, Nonmammalian Imaging, Three-Dimensional - instrumentation Imaging, Three-Dimensional - methods Life Sciences Microscopy - instrumentation Microscopy - methods Time-Lapse Imaging - instrumentation Time-Lapse Imaging - methods |
| title | Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-05T12%3A55%3A58IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_223&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Setting%20Up%20a%20Simple%20Light%20Sheet%20Microscope%20for%20In%20Toto%20Imaging%20of%20C.%20elegans%20Development&rft.jtitle=Journal%20of%20Visualized%20Experiments&rft.au=Chard%C3%A8s,%20Claire&rft.date=2014-05-05&rft.volume=87&rft.issue=87&rft.issn=1940-087X&rft.eissn=1940-087X&rft_id=info:doi/10.3791/51342&rft_dat=%3Cproquest_223%3E1526735904%3C/proquest_223%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1526735904&rft_id=info:pmid/24836407&rfr_iscdi=true |