Cadena-Herrera Daniela, Esparza-De Lara Joshua E, Ramírez-Ibañez Nancy D, López-Morales Carlos A, Pérez Néstor O, Flores-Ortiz Luis F, Medina-Rivero Emilio
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.
Biotechnol Rep (Amst). 2015 Apr 18;7:9-16. doi: 10.1016/j.btre.2015.04.004. eCollection 2015 Sep.
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.
活细胞计数对于评估细胞生长动力学至关重要。自从血细胞计数器首次用于血细胞计数以来，为了减少分析时间并通过样品制备和计数的自动化提高准确性，已经开发了几种该方法的变体。自动化技术的成功实施依赖于细胞染色、图像显示参数和细胞形态的调整，以获得与血细胞计数器相当的精密度、准确性和线性。在本研究中，我们对三种基于台盼蓝排斥的方法进行了验证：手动、半自动和全自动；这些方法用于估计用于重组蛋白生物合成和生物测定的细胞密度和活力。我们的结果表明，与手动方法相比，自动化方法的评估属性保持在相同范围内，为分析大量样品提供了一种有效的替代方法。
