Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags
Super-resolution microscopy requires that subcellular structures are labeled with bright and photostable fluorophores, especially for live-cell imaging. Organic fluorophores may help here as they can yield more photons-by orders of magnitude-than fluorescent proteins. To achieve molecular specificit...
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| Veröffentlicht in: | Cell chemical biology 2019-04, Vol.26 (4), p.584-592.e6 |
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| creator | Erdmann, Roman S Baguley, Stephanie Wood Richens, Jennifer H Wissner, Rebecca F Xi, Zhiqun Allgeyer, Edward S Zhong, Sheng Thompson, Alexander D Lowe, Nicholas Butler, Richard Bewersdorf, Joerg Rothman, James E St Johnston, Daniel Schepartz, Alanna Toomre, Derek |
| description | Super-resolution microscopy requires that subcellular structures are labeled with bright and photostable fluorophores, especially for live-cell imaging. Organic fluorophores may help here as they can yield more photons-by orders of magnitude-than fluorescent proteins. To achieve molecular specificity with organic fluorophores in live cells, self-labeling proteins are often used, with HaloTags and SNAP-tags being the most common. However, how these two different tagging systems compare with each other is unclear, especially for stimulated emission depletion (STED) microscopy, which is limited to a small repertoire of fluorophores in living cells. Herein, we compare the two labeling approaches in confocal and STED imaging using various proteins and two model systems. Strikingly, we find that the fluorescent signal can be up to 9-fold higher with HaloTags than with SNAP-tags when using far-red rhodamine derivatives. This result demonstrates that the labeling strategy matters and can greatly influence the duration of super-resolution imaging. |
| doi_str_mv | 10.1016/j.chembiol.2019.01.003 |
| format | Article |
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Organic fluorophores may help here as they can yield more photons-by orders of magnitude-than fluorescent proteins. To achieve molecular specificity with organic fluorophores in live cells, self-labeling proteins are often used, with HaloTags and SNAP-tags being the most common. However, how these two different tagging systems compare with each other is unclear, especially for stimulated emission depletion (STED) microscopy, which is limited to a small repertoire of fluorophores in living cells. Herein, we compare the two labeling approaches in confocal and STED imaging using various proteins and two model systems. Strikingly, we find that the fluorescent signal can be up to 9-fold higher with HaloTags than with SNAP-tags when using far-red rhodamine derivatives. This result demonstrates that the labeling strategy matters and can greatly influence the duration of super-resolution imaging.</description><identifier>ISSN: 2451-9456</identifier><identifier>EISSN: 2451-9448</identifier><identifier>EISSN: 2451-9456</identifier><identifier>DOI: 10.1016/j.chembiol.2019.01.003</identifier><identifier>PMID: 30745239</identifier><language>eng</language><publisher>United States: Cell Press</publisher><subject>Animals ; Drosophila ; Fluorescent Dyes - analysis ; Green Fluorescent Proteins - analysis ; HeLa Cells ; Humans ; Microscopy, Confocal - methods ; Microscopy, Fluorescence - methods ; Proteins - analysis ; Recombinant Fusion Proteins - analysis ; Rhodamines - analysis ; Staining and Labeling - methods</subject><ispartof>Cell chemical biology, 2019-04, Vol.26 (4), p.584-592.e6</ispartof><rights>Copyright © 2019 Elsevier Ltd. All rights reserved.</rights><rights>2019 The Authors 2019</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30745239$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Erdmann, Roman S</creatorcontrib><creatorcontrib>Baguley, Stephanie Wood</creatorcontrib><creatorcontrib>Richens, Jennifer H</creatorcontrib><creatorcontrib>Wissner, Rebecca F</creatorcontrib><creatorcontrib>Xi, Zhiqun</creatorcontrib><creatorcontrib>Allgeyer, Edward S</creatorcontrib><creatorcontrib>Zhong, Sheng</creatorcontrib><creatorcontrib>Thompson, Alexander D</creatorcontrib><creatorcontrib>Lowe, Nicholas</creatorcontrib><creatorcontrib>Butler, Richard</creatorcontrib><creatorcontrib>Bewersdorf, Joerg</creatorcontrib><creatorcontrib>Rothman, James E</creatorcontrib><creatorcontrib>St Johnston, Daniel</creatorcontrib><creatorcontrib>Schepartz, Alanna</creatorcontrib><creatorcontrib>Toomre, Derek</creatorcontrib><title>Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags</title><title>Cell chemical biology</title><addtitle>Cell Chem Biol</addtitle><description>Super-resolution microscopy requires that subcellular structures are labeled with bright and photostable fluorophores, especially for live-cell imaging. Organic fluorophores may help here as they can yield more photons-by orders of magnitude-than fluorescent proteins. To achieve molecular specificity with organic fluorophores in live cells, self-labeling proteins are often used, with HaloTags and SNAP-tags being the most common. However, how these two different tagging systems compare with each other is unclear, especially for stimulated emission depletion (STED) microscopy, which is limited to a small repertoire of fluorophores in living cells. Herein, we compare the two labeling approaches in confocal and STED imaging using various proteins and two model systems. Strikingly, we find that the fluorescent signal can be up to 9-fold higher with HaloTags than with SNAP-tags when using far-red rhodamine derivatives. This result demonstrates that the labeling strategy matters and can greatly influence the duration of super-resolution imaging.</description><subject>Animals</subject><subject>Drosophila</subject><subject>Fluorescent Dyes - analysis</subject><subject>Green Fluorescent Proteins - analysis</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Microscopy, Confocal - methods</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Proteins - analysis</subject><subject>Recombinant Fusion Proteins - analysis</subject><subject>Rhodamines - analysis</subject><subject>Staining and Labeling - methods</subject><issn>2451-9456</issn><issn>2451-9448</issn><issn>2451-9456</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkG1LwzAUhYMobsz9hZE_0JqXpi9-EMZQJ2wqbn4uSXbTZbRNaTJl_96CL-iney-H83DPQWhGSUwJTa8Psd5Do6yrY0ZoERMaE8LP0JglgkZFkuTnv7tIR2jq_YGQwckzyrNLNOIkSwTjxRhVK6mgtm2FN6GXASoLHq9lCNBj43q8OXbQR6_gXX0M1rV4bXXvvHbd6QbP8cI1neytHwQF4QOgxUtZu62sPJbtDm-e5i9RGK4rdGFk7WH6PSfo7f5uu1hGq-eHx8V8FXWM8xBpRY0UwIRR2ohsyKATAZSkQpGUqOF7WUBiwGjNM6OIYVLQPGU850wyJvgE3X5xu6NqYKehHWLVZdfbRvan0klb_ldauy8r916mSZbkhA6A2V_Ar_OnMv4J5S1zsg</recordid><startdate>20190418</startdate><enddate>20190418</enddate><creator>Erdmann, Roman S</creator><creator>Baguley, Stephanie Wood</creator><creator>Richens, Jennifer H</creator><creator>Wissner, Rebecca F</creator><creator>Xi, Zhiqun</creator><creator>Allgeyer, Edward S</creator><creator>Zhong, Sheng</creator><creator>Thompson, Alexander D</creator><creator>Lowe, Nicholas</creator><creator>Butler, Richard</creator><creator>Bewersdorf, Joerg</creator><creator>Rothman, James E</creator><creator>St Johnston, Daniel</creator><creator>Schepartz, Alanna</creator><creator>Toomre, Derek</creator><general>Cell Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>5PM</scope></search><sort><creationdate>20190418</creationdate><title>Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags</title><author>Erdmann, Roman S ; 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| subjects | Animals Drosophila Fluorescent Dyes - analysis Green Fluorescent Proteins - analysis HeLa Cells Humans Microscopy, Confocal - methods Microscopy, Fluorescence - methods Proteins - analysis Recombinant Fusion Proteins - analysis Rhodamines - analysis Staining and Labeling - methods |
| title | Labeling Strategies Matter for Super-Resolution Microscopy: A Comparison between HaloTags and SNAP-tags |
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