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

Biotechnology Reports

Daniela Cadena-Herrera¹  Joshua E. Esparza-De Lara²  Nancy D. Ramírez-Ibañez²  Carlos A. López-Morales²  Luis F. Flores-Ortiz²  Néstor O. Pérez²  Emilio Medina-Rivero² 

[1] Facultad de Ciencias Naturales, Universidad Icesi, Calle 18, No. 122-135, Pance, Cali, Colombia;Unidad de Investigación y Desarrollo, Probiomed S.A. de C.V., Cruce de carreteras Acatzingo-Zumpahuacán, Tenancingo, Estado de México C.P. 52400, Mexico;

关键词: Trypan blue exclusion;    Cell viability;    Cell count;   

DOI  :  10.1016/j.btre.2015.04.004

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown