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Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry

Flow cytometry is an effective and widely used tool for determination of ploidy in fish, but it is not always possible to access the fresh samples for analysis. We investigated the potential for extended storage of fish tissue with sterlet and tench as representative species of Chondrostei and Teleo...

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Autores principales: Hubálek, Martin, Flajšhans, Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359303/
https://www.ncbi.nlm.nih.gov/pubmed/33215865
http://dx.doi.org/10.1002/cyto.a.24271
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author Hubálek, Martin
Flajšhans, Martin
author_facet Hubálek, Martin
Flajšhans, Martin
author_sort Hubálek, Martin
collection PubMed
description Flow cytometry is an effective and widely used tool for determination of ploidy in fish, but it is not always possible to access the fresh samples for analysis. We investigated the potential for extended storage of fish tissue with sterlet and tench as representative species of Chondrostei and Teleostei, using blood and fin of subadult/adult specimens and tail of larvae. Thirteen procedures for extending storage, selected for rapidity and simplicity in both field and laboratory conditions, were tested for each tissue sample. Flow cytometry was applied to fresh tissue immediately after sampling and to tissue subjected to experimental protocols, always along with species‐specific standard, after 1, 5, and 10 days storage at 0–4°C or freezing at −80°C. The fluorochrome 4′,6‐diamidine‐2′‐phenylindole dihydrochloride was used with excitation/emission maximum 358/461 nm. Based on the measurability of stored samples, evaluation of directly measured coefficients of variation of their DNA peaks and the changes in fluorescence intensity compared to fresh tissue, optimal procedures for extended storage of the selected tissue types of the model species are suggested. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry.
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spelling pubmed-83593032021-08-17 Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry Hubálek, Martin Flajšhans, Martin Cytometry A Technical Note Flow cytometry is an effective and widely used tool for determination of ploidy in fish, but it is not always possible to access the fresh samples for analysis. We investigated the potential for extended storage of fish tissue with sterlet and tench as representative species of Chondrostei and Teleostei, using blood and fin of subadult/adult specimens and tail of larvae. Thirteen procedures for extending storage, selected for rapidity and simplicity in both field and laboratory conditions, were tested for each tissue sample. Flow cytometry was applied to fresh tissue immediately after sampling and to tissue subjected to experimental protocols, always along with species‐specific standard, after 1, 5, and 10 days storage at 0–4°C or freezing at −80°C. The fluorochrome 4′,6‐diamidine‐2′‐phenylindole dihydrochloride was used with excitation/emission maximum 358/461 nm. Based on the measurability of stored samples, evaluation of directly measured coefficients of variation of their DNA peaks and the changes in fluorescence intensity compared to fresh tissue, optimal procedures for extended storage of the selected tissue types of the model species are suggested. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry. John Wiley & Sons, Inc. 2020-11-30 2021-07 /pmc/articles/PMC8359303/ /pubmed/33215865 http://dx.doi.org/10.1002/cyto.a.24271 Text en © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Technical Note
Hubálek, Martin
Flajšhans, Martin
Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry
title Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry
title_full Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry
title_fullStr Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry
title_full_unstemmed Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry
title_short Simple Field Storage of Fish Samples for Measurement of DNA Content by Flow Cytometry
title_sort simple field storage of fish samples for measurement of dna content by flow cytometry
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8359303/
https://www.ncbi.nlm.nih.gov/pubmed/33215865
http://dx.doi.org/10.1002/cyto.a.24271
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