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Quantitative Cell Culture Techniques
Published in Jay L. Nadeau, Introduction to Experimental Biophysics, 2017
Sometimes it is important to know exactly how many cells are present in a liquid culture or adherent monolayer. Sometimes only the number of cells relative to a control culture or an earlier time point is what is required. At other times, it is important to distinguish the fractions of live cells versus dead, live versus apoptotic, differentiated versus undifferentiated, or labeled versus unlabeled. In all of these cases, the cells must be counted somehow. There are three general approaches to doing this: directly counting cells by eye; using a measurable parameter (absorbance, impedance) as a surrogate for cell count; or placing the cells into a specialized flow system that permits computerized cell counting. All of these methods are very commonly used, and each has its advantages and drawbacks. In this chapter, we discuss some of the most well-established ways for quantifying populations of bacteria and mammalian cells and provide some of the protocols we have developed that are designed to help sidestep the pitfalls that can occur. At the end of this chapter, you should be comfortable obtaining a bacterial IC50 growth curve; performing an end-point mammalian toxicity assay; and setting up a simple fluorescence-activated cell sorting (FACS) experiment. We also briefly introduce some of the emerging techniques, such as real-time impedance measurements for mammalian cells, imaging cytometry, and microfluidic techniques.
Microscopy cell counting and detection with fully convolutional regression networks
Published in Computer Methods in Biomechanics and Biomedical Engineering: Imaging & Visualization, 2018
Weidi Xie, J. Alison Noble, Andrew Zisserman
Counting and detecting objects in crowded images or videos is an extremely tedious and time-consuming task encountered in many real-world applications, including biology (Arteta et al. 2012, 2014, 2015; Fiaschi et al. 2012), surveillance (Chan et al. 2008; Lempitsky & Zisserman 2010) and other applications (Barinova et al. 2012). In this paper, we focus on cell counting and detection in microscopy, but the developed methodology could equally be used in other counting or detection applications. Numerous procedures in biology and medicine require cell counting and detection, for instance: a patient’s health can be inferred from the number of red blood cells and white blood cells; in clinical pathology, cell counts from images can be used for investigating hypotheses about developmental or pathological processes; and cell concentration is important in molecular biology, where it can be used to adjust the amount of chemicals to be applied in an experiment. While detection on its own, is able to determine the presence (and quantity) of an object of interest, such as cancer cells in a pathology image, furthermore, detection can be used as seeds for further segmentation or tracking.