Validation of three viable-cell counting methods: Manual, semi-automated, and automated

A viable cell count is essential to evaluate the kinetics of cell growth. Since the hemocytometer was first used for counting blood cells, several variants of the methodology have been developed towards reducing the time of analysis and improving accuracy through automation of both sample preparatio...

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Autores:
Cadena-Herrera, Daniela
Medina-Rivero, Emilio
Flores-Ortiz, Luis F.
Pérez, Néstor O.
López-Morales, Carlos A.
Ramírez-Ibañez, Nancy D.
Esparza-De Lara, Joshua E.
Tipo de recurso:
Article of investigation
Fecha de publicación:
2015
Institución:
Universidad ICESI
Repositorio:
Repositorio ICESI
Idioma:
eng
OAI Identifier:
oai:repository.icesi.edu.co:10906/79910
Acceso en línea:
http://www.sciencedirect.com/science/article/pii/S2215017X15000235
http://hdl.handle.net/10906/79910
http://dx.doi.org/10.1016/j.btre.2015.04.004
Palabra clave:
Rights
openAccess
License
https://creativecommons.org/licenses/by-nc-nd/4.0/
Description
Summary:A viable cell count is essential to evaluate the kinetics of cell growth. Since the hemocytometer was first used for counting blood cells, several variants of the methodology have been developed towards reducing the time of analysis and improving accuracy through automation of both sample preparation and counting. The successful implementation of automated techniques relies in the adjustment of cell staining, image display parameters and cell morphology to obtain equivalent precision, accuracy and linearity with respect to the hemocytometer. In this study we conducted the validation of three trypan blue exclusion-based methods: manual, semi-automated, and fully automated; which were used for the estimation of density and viability of cells employed for the biosynthesis and bioassays of recombinant proteins. Our results showed that the evaluated attributes remained within the same range for the automated methods with respect to the manual, providing an efficient alternative for analyzing a huge number of samples.