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Characterizing the three‐dimensional organization of telomeres
Background Quantitative analysis can be used in combination with fluorescence microscopy. Although the human eye is able to obtain good qualitative results, when analyzing the spatial organization of telomeres in interphase nuclei, there is a need for quantitative results based on image analysis. Me...
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| Published in: | Cytometry. Part A 2005-10, Vol.67A (2), p.144-150 |
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| container_end_page | 150 |
| container_issue | 2 |
| container_start_page | 144 |
| container_title | Cytometry. Part A |
| container_volume | 67A |
| creator | Vermolen, B. J. Garini, Y. Mai, S. Mougey, V. Fest, T. Chuang, T. C.‐Y. Chuang, A. Y.‐C. Wark, L. Young, I. T. |
| description | Background
Quantitative analysis can be used in combination with fluorescence microscopy. Although the human eye is able to obtain good qualitative results, when analyzing the spatial organization of telomeres in interphase nuclei, there is a need for quantitative results based on image analysis.
Methods
We developed a tool for analyzing three‐dimensional images of telomeres stained by fluorescence in situ hybridization in interphase nuclei with DNA counterstained with 4′,6‐diamidino‐2‐phenylindole. After deconvolution of the image, we segmented individual telomeres. From the location of the telomeres we derived a distribution parameter ρT, which indicated whether the telomeres were in a disk (ρT ≫ 1) or not (ρT ≈ 1). We sorted mouse lymphocyte nuclei and measured ρT. We also performed a bromodeoxyuridine synchronous cell sorting experiment on live cells and measured ρT at several instances.
Results
Measuring ρT for nuclei in G0/G1, S, and G2 produced 1.4 ± 0.1, 1.5 ± 0.2, and 14 ± 2, respectively, showing a significant difference between G2 and G0/G1 or S. For the bromodeoxyuridine synchronous cell sorting experiment, we found a cell cycle dependency of ρT and a correlation between ρT and an observer.
Conclusions
In this study we present a quantitative method to characterize the organization of telomeres using three‐dimensional imaging, image processing, and image analysis. © 2005 International Society for Analytical Cytology |
| doi_str_mv | 10.1002/cyto.a.20159 |
| format | article |
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Quantitative analysis can be used in combination with fluorescence microscopy. Although the human eye is able to obtain good qualitative results, when analyzing the spatial organization of telomeres in interphase nuclei, there is a need for quantitative results based on image analysis.
Methods
We developed a tool for analyzing three‐dimensional images of telomeres stained by fluorescence in situ hybridization in interphase nuclei with DNA counterstained with 4′,6‐diamidino‐2‐phenylindole. After deconvolution of the image, we segmented individual telomeres. From the location of the telomeres we derived a distribution parameter ρT, which indicated whether the telomeres were in a disk (ρT ≫ 1) or not (ρT ≈ 1). We sorted mouse lymphocyte nuclei and measured ρT. We also performed a bromodeoxyuridine synchronous cell sorting experiment on live cells and measured ρT at several instances.
Results
Measuring ρT for nuclei in G0/G1, S, and G2 produced 1.4 ± 0.1, 1.5 ± 0.2, and 14 ± 2, respectively, showing a significant difference between G2 and G0/G1 or S. For the bromodeoxyuridine synchronous cell sorting experiment, we found a cell cycle dependency of ρT and a correlation between ρT and an observer.
Conclusions
In this study we present a quantitative method to characterize the organization of telomeres using three‐dimensional imaging, image processing, and image analysis. © 2005 International Society for Analytical Cytology</description><identifier>ISSN: 1552-4922</identifier><identifier>EISSN: 1552-4930</identifier><identifier>DOI: 10.1002/cyto.a.20159</identifier><identifier>PMID: 16163697</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Algorithms ; Animals ; B-Lymphocytes - cytology ; Bromodeoxyuridine ; Cell Cycle ; Cell Nucleus ; fluorescence in situ hybridization ; fluorescence microscopy ; image processing ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional - methods ; In Situ Hybridization, Fluorescence - methods ; Mice ; Microscopy ; Telomere - chemistry ; Telomere - metabolism ; telomeres ; three‐dimensional imaging</subject><ispartof>Cytometry. Part A, 2005-10, Vol.67A (2), p.144-150</ispartof><rights>Copyright © 2005 Wiley‐Liss, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3719-e952b81ae37cc20e552966a8953c8cbf422b9f92ecf5b724ef4cb6001ac9de373</citedby><cites>FETCH-LOGICAL-c3719-e952b81ae37cc20e552966a8953c8cbf422b9f92ecf5b724ef4cb6001ac9de373</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16163697$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Vermolen, B. J.</creatorcontrib><creatorcontrib>Garini, Y.</creatorcontrib><creatorcontrib>Mai, S.</creatorcontrib><creatorcontrib>Mougey, V.</creatorcontrib><creatorcontrib>Fest, T.</creatorcontrib><creatorcontrib>Chuang, T. C.‐Y.</creatorcontrib><creatorcontrib>Chuang, A. Y.‐C.</creatorcontrib><creatorcontrib>Wark, L.</creatorcontrib><creatorcontrib>Young, I. T.</creatorcontrib><title>Characterizing the three‐dimensional organization of telomeres</title><title>Cytometry. Part A</title><addtitle>Cytometry A</addtitle><description>Background
Quantitative analysis can be used in combination with fluorescence microscopy. Although the human eye is able to obtain good qualitative results, when analyzing the spatial organization of telomeres in interphase nuclei, there is a need for quantitative results based on image analysis.
Methods
We developed a tool for analyzing three‐dimensional images of telomeres stained by fluorescence in situ hybridization in interphase nuclei with DNA counterstained with 4′,6‐diamidino‐2‐phenylindole. After deconvolution of the image, we segmented individual telomeres. From the location of the telomeres we derived a distribution parameter ρT, which indicated whether the telomeres were in a disk (ρT ≫ 1) or not (ρT ≈ 1). We sorted mouse lymphocyte nuclei and measured ρT. We also performed a bromodeoxyuridine synchronous cell sorting experiment on live cells and measured ρT at several instances.
Results
Measuring ρT for nuclei in G0/G1, S, and G2 produced 1.4 ± 0.1, 1.5 ± 0.2, and 14 ± 2, respectively, showing a significant difference between G2 and G0/G1 or S. For the bromodeoxyuridine synchronous cell sorting experiment, we found a cell cycle dependency of ρT and a correlation between ρT and an observer.
Conclusions
In this study we present a quantitative method to characterize the organization of telomeres using three‐dimensional imaging, image processing, and image analysis. © 2005 International Society for Analytical Cytology</description><subject>Algorithms</subject><subject>Animals</subject><subject>B-Lymphocytes - cytology</subject><subject>Bromodeoxyuridine</subject><subject>Cell Cycle</subject><subject>Cell Nucleus</subject><subject>fluorescence in situ hybridization</subject><subject>fluorescence microscopy</subject><subject>image processing</subject><subject>Image Processing, Computer-Assisted</subject><subject>Imaging, Three-Dimensional - methods</subject><subject>In Situ Hybridization, Fluorescence - methods</subject><subject>Mice</subject><subject>Microscopy</subject><subject>Telomere - chemistry</subject><subject>Telomere - metabolism</subject><subject>telomeres</subject><subject>three‐dimensional imaging</subject><issn>1552-4922</issn><issn>1552-4930</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><recordid>eNp9kD1PwzAQhi0EoqWwMaNMTKT4I3biDRTxJVXqUgYmy3EvrVESFzsVaid-Ar-RX0JKKtgYTncnPffo9CJ0TvCYYEyvzaZ1Yz2mmHB5gIaEcxonkuHD35nSAToJ4RVjxjGjx2hABBFMyHSIbvKl9tq04O3WNouoXUJXHuDr43Nua2iCdY2uIucXurFb3XZr5MqohcrV4CGcoqNSVwHO9n2Enu_vZvljPJk-POW3k9iwlMgYJKdFRjSw1BiKoftMCqEzyZnJTFEmlBaylBRMyYuUJlAmphAYE23kvDtiI3TZe1feva0htKq2wUBV6QbcOiiRCULTDHfgVQ8a70LwUKqVt7X2G0Ww2iWmdokprX4S6_CLvXdd1DD_g_cRdQDrgXdbweZfmcpfZtNe-w1KUXmw</recordid><startdate>200510</startdate><enddate>200510</enddate><creator>Vermolen, B. J.</creator><creator>Garini, Y.</creator><creator>Mai, S.</creator><creator>Mougey, V.</creator><creator>Fest, T.</creator><creator>Chuang, T. C.‐Y.</creator><creator>Chuang, A. Y.‐C.</creator><creator>Wark, L.</creator><creator>Young, I. T.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><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></search><sort><creationdate>200510</creationdate><title>Characterizing the three‐dimensional organization of telomeres</title><author>Vermolen, B. J. ; Garini, Y. ; Mai, S. ; Mougey, V. ; Fest, T. ; Chuang, T. C.‐Y. ; Chuang, A. Y.‐C. ; Wark, L. ; Young, I. T.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3719-e952b81ae37cc20e552966a8953c8cbf422b9f92ecf5b724ef4cb6001ac9de373</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Algorithms</topic><topic>Animals</topic><topic>B-Lymphocytes - cytology</topic><topic>Bromodeoxyuridine</topic><topic>Cell Cycle</topic><topic>Cell Nucleus</topic><topic>fluorescence in situ hybridization</topic><topic>fluorescence microscopy</topic><topic>image processing</topic><topic>Image Processing, Computer-Assisted</topic><topic>Imaging, Three-Dimensional - methods</topic><topic>In Situ Hybridization, Fluorescence - methods</topic><topic>Mice</topic><topic>Microscopy</topic><topic>Telomere - chemistry</topic><topic>Telomere - metabolism</topic><topic>telomeres</topic><topic>three‐dimensional imaging</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Vermolen, B. J.</creatorcontrib><creatorcontrib>Garini, Y.</creatorcontrib><creatorcontrib>Mai, S.</creatorcontrib><creatorcontrib>Mougey, V.</creatorcontrib><creatorcontrib>Fest, T.</creatorcontrib><creatorcontrib>Chuang, T. C.‐Y.</creatorcontrib><creatorcontrib>Chuang, A. Y.‐C.</creatorcontrib><creatorcontrib>Wark, L.</creatorcontrib><creatorcontrib>Young, I. T.</creatorcontrib><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><jtitle>Cytometry. Part A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Vermolen, B. J.</au><au>Garini, Y.</au><au>Mai, S.</au><au>Mougey, V.</au><au>Fest, T.</au><au>Chuang, T. C.‐Y.</au><au>Chuang, A. Y.‐C.</au><au>Wark, L.</au><au>Young, I. T.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterizing the three‐dimensional organization of telomeres</atitle><jtitle>Cytometry. Part A</jtitle><addtitle>Cytometry A</addtitle><date>2005-10</date><risdate>2005</risdate><volume>67A</volume><issue>2</issue><spage>144</spage><epage>150</epage><pages>144-150</pages><issn>1552-4922</issn><eissn>1552-4930</eissn><abstract>Background
Quantitative analysis can be used in combination with fluorescence microscopy. Although the human eye is able to obtain good qualitative results, when analyzing the spatial organization of telomeres in interphase nuclei, there is a need for quantitative results based on image analysis.
Methods
We developed a tool for analyzing three‐dimensional images of telomeres stained by fluorescence in situ hybridization in interphase nuclei with DNA counterstained with 4′,6‐diamidino‐2‐phenylindole. After deconvolution of the image, we segmented individual telomeres. From the location of the telomeres we derived a distribution parameter ρT, which indicated whether the telomeres were in a disk (ρT ≫ 1) or not (ρT ≈ 1). We sorted mouse lymphocyte nuclei and measured ρT. We also performed a bromodeoxyuridine synchronous cell sorting experiment on live cells and measured ρT at several instances.
Results
Measuring ρT for nuclei in G0/G1, S, and G2 produced 1.4 ± 0.1, 1.5 ± 0.2, and 14 ± 2, respectively, showing a significant difference between G2 and G0/G1 or S. For the bromodeoxyuridine synchronous cell sorting experiment, we found a cell cycle dependency of ρT and a correlation between ρT and an observer.
Conclusions
In this study we present a quantitative method to characterize the organization of telomeres using three‐dimensional imaging, image processing, and image analysis. © 2005 International Society for Analytical Cytology</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>16163697</pmid><doi>10.1002/cyto.a.20159</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Cytometry. Part A, 2005-10, Vol.67A (2), p.144-150 |
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| source | Wiley-Blackwell Read & Publish Collection |
| subjects | Algorithms Animals B-Lymphocytes - cytology Bromodeoxyuridine Cell Cycle Cell Nucleus fluorescence in situ hybridization fluorescence microscopy image processing Image Processing, Computer-Assisted Imaging, Three-Dimensional - methods In Situ Hybridization, Fluorescence - methods Mice Microscopy Telomere - chemistry Telomere - metabolism telomeres three‐dimensional imaging |
| title | Characterizing the three‐dimensional organization of telomeres |
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