Measuring the Bending Stiffness of Bacterial Cells Using an Optical Trap
We developed a protocol to measure the bending rigidity of filamentous rod-shaped bacteria. Forces are applied with an optical trap, a microscopic three-dimensional spring made of light that is formed when a high-intensity laser beam is focused to a very small spot by a microscope's objective l...
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| Veröffentlicht in: | Journal of Visualized Experiments 2010-04 (38) |
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| creator | Wang, Siyuan Arellano-Santoyo, Hugo Combs, Peter A. Shaevitz, Joshua W. |
| description | We developed a protocol to measure the bending rigidity of filamentous rod-shaped bacteria. Forces are applied with an optical trap, a microscopic three-dimensional spring made of light that is formed when a high-intensity laser beam is focused to a very small spot by a microscope's objective lens. To bend a cell, we first bind live bacteria to a chemically-treated coverslip. As these cells grow, the middle of the cells remains bound to the coverslip but the growing ends are free of this restraint. By inducing filamentous growth with the drug cephalexin, we are able to identify cells in which one end of the cell was stuck to the surface while the other end remained unattached and susceptible to bending forces. A bending force is then applied with an optical trap by binding a polylysine-coated bead to the tip of a growing cell. Both the force and the displacement of the bead are recorded and the bending stiffness of the cell is the slope of this relationship. |
| doi_str_mv | 10.3791/2012 |
| format | Article |
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Forces are applied with an optical trap, a microscopic three-dimensional spring made of light that is formed when a high-intensity laser beam is focused to a very small spot by a microscope's objective lens. To bend a cell, we first bind live bacteria to a chemically-treated coverslip. As these cells grow, the middle of the cells remains bound to the coverslip but the growing ends are free of this restraint. By inducing filamentous growth with the drug cephalexin, we are able to identify cells in which one end of the cell was stuck to the surface while the other end remained unattached and susceptible to bending forces. A bending force is then applied with an optical trap by binding a polylysine-coated bead to the tip of a growing cell. Both the force and the displacement of the bead are recorded and the bending stiffness of the cell is the slope of this relationship.</description><subject>Bacteriological Techniques - methods</subject><subject>Cephalexin - pharmacology</subject><subject>Escherichia coli - drug effects</subject><subject>Escherichia coli - growth & development</subject><subject>Escherichia coli - physiology</subject><subject>Microbiology</subject><subject>Optical Tweezers</subject><subject>Polylysine - chemistry</subject><issn>1940-087X</issn><issn>1940-087X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkMFOwzAMhiMEYmPjAbigHpA4FZImTZsLEpuAIQ3twCZxi9LU3Tp1TUnaSbw9rTqmcbJlf_5t_wiNCX6gkSCPASbBGRoSwbCP4-jr_CQfoCvnthjzAIfxJRoEmAUk5myIZh-gXGPzcu3VG_AmUKZd_lnnWVaCc57JvInSNdhcFd4UisJ5K9chqvQWVZ3rtry0qhqji0wVDq4PcYRWry_L6cyfL97ep89zX1MeBD5lHDIdJXHIeCyYSEQSAedhGoVAUkopjwEHScpSwDHGEZAsFSJNNNYqTERGR-ip162aZAephrK2qpCVzXfK_kijcvm_U-YbuTZ7SQlnOCatwP1BwJrvBlwtd7nT7WOqBNM4GVEqCAtD3pJ3Pamtcc5CdtxCsOw8l53nLXZ7etER-jO5BW56YGv2ILemsWXrUD_8C7nhhAk</recordid><startdate>20100426</startdate><enddate>20100426</enddate><creator>Wang, Siyuan</creator><creator>Arellano-Santoyo, Hugo</creator><creator>Combs, Peter A.</creator><creator>Shaevitz, Joshua W.</creator><general>MyJove Corporation</general><scope>ALOKQ</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>5PM</scope></search><sort><creationdate>20100426</creationdate><title>Measuring the Bending Stiffness of Bacterial Cells Using an Optical Trap</title><author>Wang, Siyuan ; Arellano-Santoyo, Hugo ; Combs, Peter A. ; Shaevitz, Joshua W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3622-346efc7b85468949b9b7e665d75e1d33368e02bd4de08007e1fd99dbc0ca5b9f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Bacteriological Techniques - methods</topic><topic>Cephalexin - pharmacology</topic><topic>Escherichia coli - drug effects</topic><topic>Escherichia coli - growth & development</topic><topic>Escherichia coli - physiology</topic><topic>Microbiology</topic><topic>Optical Tweezers</topic><topic>Polylysine - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Siyuan</creatorcontrib><creatorcontrib>Arellano-Santoyo, Hugo</creatorcontrib><creatorcontrib>Combs, Peter A.</creatorcontrib><creatorcontrib>Shaevitz, Joshua W.</creatorcontrib><collection>JoVE Journal: Biology</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>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>Wang, Siyuan</au><au>Arellano-Santoyo, Hugo</au><au>Combs, Peter A.</au><au>Shaevitz, Joshua W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Measuring the Bending Stiffness of Bacterial Cells Using an Optical Trap</atitle><jtitle>Journal of Visualized Experiments</jtitle><addtitle>J Vis Exp</addtitle><date>2010-04-26</date><risdate>2010</risdate><issue>38</issue><issn>1940-087X</issn><eissn>1940-087X</eissn><abstract>We developed a protocol to measure the bending rigidity of filamentous rod-shaped bacteria. Forces are applied with an optical trap, a microscopic three-dimensional spring made of light that is formed when a high-intensity laser beam is focused to a very small spot by a microscope's objective lens. To bend a cell, we first bind live bacteria to a chemically-treated coverslip. As these cells grow, the middle of the cells remains bound to the coverslip but the growing ends are free of this restraint. By inducing filamentous growth with the drug cephalexin, we are able to identify cells in which one end of the cell was stuck to the surface while the other end remained unattached and susceptible to bending forces. A bending force is then applied with an optical trap by binding a polylysine-coated bead to the tip of a growing cell. Both the force and the displacement of the bead are recorded and the bending stiffness of the cell is the slope of this relationship.</abstract><cop>United States</cop><pub>MyJove Corporation</pub><pmid>20421864</pmid><doi>10.3791/2012</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Bacteriological Techniques - methods Cephalexin - pharmacology Escherichia coli - drug effects Escherichia coli - growth & development Escherichia coli - physiology Microbiology Optical Tweezers Polylysine - chemistry |
| title | Measuring the Bending Stiffness of Bacterial Cells Using an Optical Trap |
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