Sensitive detection of fluorescence in western blotting by merging images
The western blotting technique is widely used to analyze protein expression levels and protein molecular weight. The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to det...
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| Veröffentlicht in: | PloS one 2018-01, Vol.13 (1), p.e0191532-e0191532 |
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| creator | Kondo, Yukari Higa, Shinichiro Iwasaki, Takeshi Matsumoto, Tomoya Maehara, Kazumitsu Harada, Akihito Baba, Yoshihiro Fujita, Masatoshi Ohkawa, Yasuyuki |
| description | The western blotting technique is widely used to analyze protein expression levels and protein molecular weight. The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously. Recently, more attention has been paid to the fluorescence detection method because it is more quantitative and is suitable for the detection of multiple proteins simultaneously. However, fluorescence detection can be limited by poor image resolution and low detection sensitivity. Here, we describe a method to detect fluorescence in western blots using fluorescence microscopy to obtain high-resolution images. In this method, filters and fluorescent dyes are optimized to enhance detection sensitivity to a level similar to that of the chemiluminescence method. |
| doi_str_mv | 10.1371/journal.pone.0191532 |
| format | Article |
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The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously. Recently, more attention has been paid to the fluorescence detection method because it is more quantitative and is suitable for the detection of multiple proteins simultaneously. However, fluorescence detection can be limited by poor image resolution and low detection sensitivity. Here, we describe a method to detect fluorescence in western blots using fluorescence microscopy to obtain high-resolution images. In this method, filters and fluorescent dyes are optimized to enhance detection sensitivity to a level similar to that of the chemiluminescence method.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0191532</identifier><identifier>PMID: 29352284</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Biochemistry ; Biology and Life Sciences ; Chemiluminescence ; Engineering and Technology ; Fluorescence ; Fluorescence microscopy ; Fluorescent dyes ; Fluorescent indicators ; Image detection ; Image enhancement ; Image filters ; Image processing ; Image resolution ; Immunoglobulins ; Life sciences ; Measurement ; Methods ; Microscopy ; Molecular weight ; Noise ; Pharmaceutical sciences ; Physical Sciences ; Protein expression ; Proteins ; Research and Analysis Methods ; Sensitivity enhancement ; Western blotting</subject><ispartof>PloS one, 2018-01, Vol.13 (1), p.e0191532-e0191532</ispartof><rights>COPYRIGHT 2018 Public Library of Science</rights><rights>2018 Kondo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2018 Kondo et al 2018 Kondo et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-7f8bb98bb3e522b5495392f697caca4c8d7604763c9824e338af8f5e7a5e6bf23</citedby><cites>FETCH-LOGICAL-c692t-7f8bb98bb3e522b5495392f697caca4c8d7604763c9824e338af8f5e7a5e6bf23</cites><orcidid>0000-0001-6440-9954</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5774814/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5774814/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29352284$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Asakura, Atsushi</contributor><creatorcontrib>Kondo, Yukari</creatorcontrib><creatorcontrib>Higa, Shinichiro</creatorcontrib><creatorcontrib>Iwasaki, Takeshi</creatorcontrib><creatorcontrib>Matsumoto, Tomoya</creatorcontrib><creatorcontrib>Maehara, Kazumitsu</creatorcontrib><creatorcontrib>Harada, Akihito</creatorcontrib><creatorcontrib>Baba, Yoshihiro</creatorcontrib><creatorcontrib>Fujita, Masatoshi</creatorcontrib><creatorcontrib>Ohkawa, Yasuyuki</creatorcontrib><title>Sensitive detection of fluorescence in western blotting by merging images</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The western blotting technique is widely used to analyze protein expression levels and protein molecular weight. The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously. Recently, more attention has been paid to the fluorescence detection method because it is more quantitative and is suitable for the detection of multiple proteins simultaneously. However, fluorescence detection can be limited by poor image resolution and low detection sensitivity. Here, we describe a method to detect fluorescence in western blots using fluorescence microscopy to obtain high-resolution images. 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The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously. Recently, more attention has been paid to the fluorescence detection method because it is more quantitative and is suitable for the detection of multiple proteins simultaneously. However, fluorescence detection can be limited by poor image resolution and low detection sensitivity. Here, we describe a method to detect fluorescence in western blots using fluorescence microscopy to obtain high-resolution images. In this method, filters and fluorescent dyes are optimized to enhance detection sensitivity to a level similar to that of the chemiluminescence method.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>29352284</pmid><doi>10.1371/journal.pone.0191532</doi><tpages>e0191532</tpages><orcidid>https://orcid.org/0000-0001-6440-9954</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Biochemistry Biology and Life Sciences Chemiluminescence Engineering and Technology Fluorescence Fluorescence microscopy Fluorescent dyes Fluorescent indicators Image detection Image enhancement Image filters Image processing Image resolution Immunoglobulins Life sciences Measurement Methods Microscopy Molecular weight Noise Pharmaceutical sciences Physical Sciences Protein expression Proteins Research and Analysis Methods Sensitivity enhancement Western blotting |
| title | Sensitive detection of fluorescence in western blotting by merging images |
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