Lawrence Livermore National Laboratory, Livermore, California 94550, USA.
Biotechnol Bioeng. 2013 Mar;110(3):838-47. doi: 10.1002/bit.24757. Epub 2013 Jan 7.
We have developed an automated cell counting method that uses images obtained at multiple focal heights to enumerate cells in confluent culture. By taking the derivative of image intensity with respect to focal height using two complementary images, we are able to count high-density monolayers of cells over a large image area. Our method resists errors arising from variability in the focal plane caused by flatness or tilt non-uniformities with a minimal amount of focal plane alignment, allowing the automated collection of images across a large area.
我们开发了一种自动细胞计数方法，该方法使用在多个焦平面获得的图像来对汇合培养中的细胞进行计数。通过使用两个互补图像对图像强度相对于焦平面的导数进行计算，我们能够对大图像区域中的高密度单层细胞进行计数。我们的方法通过最小量的焦平面对准来抵抗由于平整度或倾斜不均匀性引起的焦平面变化引起的误差，从而允许在大面积上自动采集图像。
