Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.cryobiol.2020.06.004 http://hdl.handle.net/11449/197152 |
Resumo: | Flow cytometry is a valuable tool in biomedical and animal sciences. However, equipment used for such analysis presents limitations at field conditions, suggesting then preservation procedures for future analysis at laboratory conditions. In this study, freezing at low (-20 degrees C), ultra-low (-80 degrees C) and cryogenic temperatures (-196 degrees C, i.e. liquid nitrogen) were used as preservation procedures of fish tissue. Samples were maintained in 0.9% NaCl or lysing solution, and stored at the temperatures above for 0 (fresh control), 60, 120 and 180 days of storage. After storage, the samples were thawed and proceeded to flow cytometric analysis. Storage at low temperatures (-20 degrees C), both in lysing and 0.9% NaCl, exhibited poor results when analyzed after 60, 120 and 180 days, showing noisy peaks, deviation in the DNA content and absence of peaks. Ultralow (-80 degrees C) and cryogenic (-196 degrees C) temperatures, both in lysing solution and 0.9% NaCl, showed good results and high quality of histograms. Both storage procedures gave similar histograms and DNA content in comparison with control group (fresh) even after 60, 120 and 180 days of storage, exhibiting the main peak at 2C content from diploid cells and a secondary peak at 4C derived from dividing cells. In conclusion, samples may be stored for 180 days at -80 degrees C and -196 degrees C in both, 0.9% NaCl or lysing solution. As cryogenic temperatures in liquid nitrogen permits indefinite storage, this procedure should be used for long-term preservation. |
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Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperaturesCell cycleDNA quantityFishPloidyStorageFlow cytometry is a valuable tool in biomedical and animal sciences. However, equipment used for such analysis presents limitations at field conditions, suggesting then preservation procedures for future analysis at laboratory conditions. In this study, freezing at low (-20 degrees C), ultra-low (-80 degrees C) and cryogenic temperatures (-196 degrees C, i.e. liquid nitrogen) were used as preservation procedures of fish tissue. Samples were maintained in 0.9% NaCl or lysing solution, and stored at the temperatures above for 0 (fresh control), 60, 120 and 180 days of storage. After storage, the samples were thawed and proceeded to flow cytometric analysis. Storage at low temperatures (-20 degrees C), both in lysing and 0.9% NaCl, exhibited poor results when analyzed after 60, 120 and 180 days, showing noisy peaks, deviation in the DNA content and absence of peaks. Ultralow (-80 degrees C) and cryogenic (-196 degrees C) temperatures, both in lysing solution and 0.9% NaCl, showed good results and high quality of histograms. Both storage procedures gave similar histograms and DNA content in comparison with control group (fresh) even after 60, 120 and 180 days of storage, exhibiting the main peak at 2C content from diploid cells and a secondary peak at 4C derived from dividing cells. In conclusion, samples may be stored for 180 days at -80 degrees C and -196 degrees C in both, 0.9% NaCl or lysing solution. As cryogenic temperatures in liquid nitrogen permits indefinite storage, this procedure should be used for long-term preservation.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)AES Tiete (Research & Development Program ANEEL)Univ Sao Paulo, Dept Anim Reprod FMVZ, Ave Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP, BrazilChico Mendes Inst Biodivers Conservat, Lab Fish Biotechnol, Natl Ctr Res & Conservat Continental Fish, Rodovia Pref Euberto Nemesio Pereira de Godoy, BR-13630970 Pirassununga, SP, BrazilSao Paulo State Univ Julio de Mesquita Filho, Inst Biosci, Rua Prof Dr Antonio Celso Wagner Zanin 250, BR-18618689 Botucatu, SP, BrazilUniv Sao Paulo, Dept Vet Med FZEA, Ave Duque Caxias Norte 225, BR-13630080 Pirassununga, SP, BrazilSao Paulo State Univ, Aquaculture Ctr, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, BrazilAES Tiete, Br-153,0 Km 139 Ctr, BR-16370000 Promissao, SP, BrazilSao Paulo State Univ Julio de Mesquita Filho, Inst Biosci, Rua Prof Dr Antonio Celso Wagner Zanin 250, BR-18618689 Botucatu, SP, BrazilSao Paulo State Univ, Aquaculture Ctr, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, BrazilFAPESP: 2010/17429-1FAPESP: 2011/11664-1AES Tiete (Research & Development Program ANEEL): 0064-1042/2014Elsevier B.V.Universidade de São Paulo (USP)Chico Mendes Inst Biodivers ConservatUniversidade Estadual Paulista (Unesp)AES TieteYasui, George ShiguekiBertolini, Rafaela Manchin [UNESP]Suarez-Lopez, Lucia [UNESP]Xavier, Pedro PorfirioMonzani, Paulo SergioNascimento, Nivaldo Ferreira doCastilho, Antonio Leao [UNESP]Okada Nakaghi, Laura Satiko [UNESP]Alves dos Santos, Silvio CarlosSenhorini, Jose Augusto2020-12-10T20:07:47Z2020-12-10T20:07:47Z2020-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article68-71http://dx.doi.org/10.1016/j.cryobiol.2020.06.004Cryobiology. San Diego: Academic Press Inc Elsevier Science, v. 95, p. 68-71, 2020.0011-2240http://hdl.handle.net/11449/19715210.1016/j.cryobiol.2020.06.004WOS:000556580400025Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengCryobiologyinfo:eu-repo/semantics/openAccess2024-04-09T15:10:26Zoai:repositorio.unesp.br:11449/197152Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T13:46:15.282519Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
title |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
spellingShingle |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures Yasui, George Shigueki Cell cycle DNA quantity Fish Ploidy Storage |
title_short |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
title_full |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
title_fullStr |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
title_full_unstemmed |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
title_sort |
Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures |
author |
Yasui, George Shigueki |
author_facet |
Yasui, George Shigueki Bertolini, Rafaela Manchin [UNESP] Suarez-Lopez, Lucia [UNESP] Xavier, Pedro Porfirio Monzani, Paulo Sergio Nascimento, Nivaldo Ferreira do Castilho, Antonio Leao [UNESP] Okada Nakaghi, Laura Satiko [UNESP] Alves dos Santos, Silvio Carlos Senhorini, Jose Augusto |
author_role |
author |
author2 |
Bertolini, Rafaela Manchin [UNESP] Suarez-Lopez, Lucia [UNESP] Xavier, Pedro Porfirio Monzani, Paulo Sergio Nascimento, Nivaldo Ferreira do Castilho, Antonio Leao [UNESP] Okada Nakaghi, Laura Satiko [UNESP] Alves dos Santos, Silvio Carlos Senhorini, Jose Augusto |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Chico Mendes Inst Biodivers Conservat Universidade Estadual Paulista (Unesp) AES Tiete |
dc.contributor.author.fl_str_mv |
Yasui, George Shigueki Bertolini, Rafaela Manchin [UNESP] Suarez-Lopez, Lucia [UNESP] Xavier, Pedro Porfirio Monzani, Paulo Sergio Nascimento, Nivaldo Ferreira do Castilho, Antonio Leao [UNESP] Okada Nakaghi, Laura Satiko [UNESP] Alves dos Santos, Silvio Carlos Senhorini, Jose Augusto |
dc.subject.por.fl_str_mv |
Cell cycle DNA quantity Fish Ploidy Storage |
topic |
Cell cycle DNA quantity Fish Ploidy Storage |
description |
Flow cytometry is a valuable tool in biomedical and animal sciences. However, equipment used for such analysis presents limitations at field conditions, suggesting then preservation procedures for future analysis at laboratory conditions. In this study, freezing at low (-20 degrees C), ultra-low (-80 degrees C) and cryogenic temperatures (-196 degrees C, i.e. liquid nitrogen) were used as preservation procedures of fish tissue. Samples were maintained in 0.9% NaCl or lysing solution, and stored at the temperatures above for 0 (fresh control), 60, 120 and 180 days of storage. After storage, the samples were thawed and proceeded to flow cytometric analysis. Storage at low temperatures (-20 degrees C), both in lysing and 0.9% NaCl, exhibited poor results when analyzed after 60, 120 and 180 days, showing noisy peaks, deviation in the DNA content and absence of peaks. Ultralow (-80 degrees C) and cryogenic (-196 degrees C) temperatures, both in lysing solution and 0.9% NaCl, showed good results and high quality of histograms. Both storage procedures gave similar histograms and DNA content in comparison with control group (fresh) even after 60, 120 and 180 days of storage, exhibiting the main peak at 2C content from diploid cells and a secondary peak at 4C derived from dividing cells. In conclusion, samples may be stored for 180 days at -80 degrees C and -196 degrees C in both, 0.9% NaCl or lysing solution. As cryogenic temperatures in liquid nitrogen permits indefinite storage, this procedure should be used for long-term preservation. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-12-10T20:07:47Z 2020-12-10T20:07:47Z 2020-08-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.cryobiol.2020.06.004 Cryobiology. San Diego: Academic Press Inc Elsevier Science, v. 95, p. 68-71, 2020. 0011-2240 http://hdl.handle.net/11449/197152 10.1016/j.cryobiol.2020.06.004 WOS:000556580400025 |
url |
http://dx.doi.org/10.1016/j.cryobiol.2020.06.004 http://hdl.handle.net/11449/197152 |
identifier_str_mv |
Cryobiology. San Diego: Academic Press Inc Elsevier Science, v. 95, p. 68-71, 2020. 0011-2240 10.1016/j.cryobiol.2020.06.004 WOS:000556580400025 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Cryobiology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
68-71 |
dc.publisher.none.fl_str_mv |
Elsevier B.V. |
publisher.none.fl_str_mv |
Elsevier B.V. |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808128274371444736 |