Close Dan, Xu Tingting, Ripp Steven, Sayler Gary
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, USA.
Methods Mol Biol. 2014;1098:107-16. doi: 10.1007/978-1-62703-718-1_9.
Cellular population dynamics are routinely monitored across many diverse fields for a variety of purposes. In general, these dynamics are assayed either through the direct counting of cellular aliquots followed by extrapolation to the total population size, or through the monitoring of signal intensity from any number of externally stimulated reporter proteins. While both viable methods, here we describe a novel technique that allows for the automated, non-destructive tracking of cellular population dynamics in real-time. This method, which relies on the detection of a continuous bioluminescent signal produced through expression of the bacterial luciferase gene cassette, provides a low cost, low time-intensive means for generating additional data compared to alternative methods.
出于各种目的,细胞群体动态在许多不同领域都有常规监测。一般来说,这些动态通过对细胞等分试样进行直接计数,然后外推至总群体大小来测定,或者通过监测来自任意数量外部刺激的报告蛋白的信号强度来测定。虽然这两种方法都是可行的,但在此我们描述一种新技术,它能够实时自动、无损地追踪细胞群体动态。这种方法依赖于检测通过细菌荧光素酶基因盒表达产生的连续生物发光信号,与其他方法相比,它提供了一种低成本、低时间投入的方式来生成额外数据。