Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures

Autor(es)

Yasui, George Shigueki ^{[1, 2]} ; Bertolini, Rafaela Manchin ^{[1, 3]} ; Suarez-Lopez, Lucia ^{[1, 3]} ; Xavier, Pedro Porfirio ^[4] ; Monzani, Paulo Sergio ^[1] ; do Nascimento, Nivaldo Ferreira ^{[1, 2]} ; Castilho, Antonio Leao ^[3] ; Okada Nakaghi, Laura Satiko ^[5] ; Alves dos Santos, Silvio Carlos ^[6] ; Senhorini, Jose Augusto ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Chico Mendes Inst Biodivers Conservat, Lab Fish Biotechnol, Natl Ctr Res & Conservat Continental Fish, Rodovia Pref Euberto Nemesio Pereira de Godoy, BR-13630970 Pirassununga, SP - Brazil

^[2] Univ Sao Paulo, Dept Anim Reprod FMVZ, Ave Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP - Brazil

^[3] Sao Paulo State Univ Julio de Mesquita Filho, Inst Biosci, Rua Prof Dr Antonio Celso Wagner Zanin 250, BR-18618689 Botucatu, SP - Brazil

^[4] Univ Sao Paulo, Dept Vet Med FZEA, Ave Duque Caxias Norte 225, BR-13630080 Pirassununga, SP - Brazil

^[5] Sao Paulo State Univ, Aquaculture Ctr, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP - Brazil

^[6] AES Tiete, Br-153, 0 Km 139 Ctr, BR-16370000 Promissao, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: CRYOBIOLOGY; v. 95, p. 68-71, AUG 2020.
• Citações Web of Science: 0

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. (AU)

• Processo FAPESP: 10/17429-1 - Quimerismo em espécies nativas de peixes como ferramenta para a propagação mediada na aquicultura
• Beneficiário: George Shigueki Yasui
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 11/11664-1 - Quimerismo em espécies nativas de peixes como ferramenta para a propagação mediada na aquicultura
• Beneficiário: George Shigueki Yasui
• Modalidade de apoio: Bolsas no Brasil - Jovens Pesquisadores