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Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures.
Metadata
Journalcryobiology2.283Date
2020 Jun 05
4 months ago
Type
Journal Article
Volume
2020-Aug / 95 : 68-71
Author
Yasui GS 1, Bertolini RM 2, Suárez-López L 2, Xavier PP 3, Monzani PS 4, Ferreira do Nascimento N 5, Castilho AL 6, Okada Nakaghi LS 7, Alves Dos Santos SC 8, Senhorini JA 5
Affiliation
  • 2. Laboratory of Fish Biotechnology, National Center for Research and Conservation of Continental Fish, Chico Mendes Institute of Biodiversity Conservation, Rodovia Pref. Euberto Nemésio Pereira de Godoy, Pirassununga, SP, 13630-970, Brazil; Institute of Biosciences, São Paulo State University Júlio de Mesquita Filho, Rua Prof. Dr. Antônio Celso Wagner Zanin 250, Rubião Junior District, 18618-689, São Paulo, Botucatu, Brazil.
  • 3. Department of Veterinary Medicine - FZEA, University of Sao Paulo, Avenida Duque de Caxias Norte 225, Pirassununga, SP, 13630-080, Brazil.
  • 4. Laboratory of Fish Biotechnology, National Center for Research and Conservation of Continental Fish, Chico Mendes Institute of Biodiversity Conservation, Rodovia Pref. Euberto Nemésio Pereira de Godoy, Pirassununga, SP, 13630-970, Brazil.
  • 5. Department of Animal Reproduction - FMVZ, University of Sao Paulo, Avenida Prof. Dr. Orlando Marques de Paiva, 87, São Paulo-SP, 05508-270, Brazil; Laboratory of Fish Biotechnology, National Center for Research and Conservation of Continental Fish, Chico Mendes Institute of Biodiversity Conservation, Rodovia Pref. Euberto Nemésio Pereira de Godoy, Pirassununga, SP, 13630-970, Brazil.
  • 6. Institute of Biosciences, São Paulo State University Júlio de Mesquita Filho, Rua Prof. Dr. Antônio Celso Wagner Zanin 250, Rubião Junior District, 18618-689, São Paulo, Botucatu, Brazil.
  • 7. Aquaculture Center, Sao Paulo State University, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP, 14884-900, Brazil.
  • 8. AES Tietê, Br-153, Rod, 0 Km 139 Centro, Promissão, SP, 16370-000, Brazil.
Doi
PMIDMESH
Abstract
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 °C), ultra-low (-80 °C) and cryogenic temperatures (-196 °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 °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 °C) and cryogenic (-196 °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 °C and -196 °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.
Keywords: Cell cycle DNA quantity Fish Ploidy Storage
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Cryobiologycryobiology
Metadata
LocationNetherlands
FromACADEMIC PRESS INC ELSEVIER SCIENCE

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