Flow cytometers, on the other hand, are expensive but they have many applications such as cell counting, analysis of the cells shape, structure and measuring internal cell markers 4,5. However, they are rarely used for only cell counting due to their considerable cost compared to manual counting 3. They are fast, accurate and cheaper than flow cytometers. Coulter counters take advantage of cells electrical resistance to determine cell number and size. Most of the automated methods for cell counting belong to one of two classes, those that use the Coulter principle or flow cytometry. Cell counting can be performed manually using a hemocytometer as well as using a variety of automated methods, each with their own advantages and disadvantages 3,4,5. Accurately determining the number of cells in a culture is essential for experimental reproducibility and standardization 1,2. In vitro cell counting is an important basic technique in a wide range of tissue culture experiments. By combining the core principles of Cell Counter with an automated counting algorithm and post-counting analysis, this greatly increases the ease with which migration assays can be processed without any loss of accuracy. They are easy to use and optimized for quick counting and analysis of large sample sizes with built-in analysis tools to help calibration of counts. Both plugins rely on the ability to acquire high quality micrographs with minimal background. To address this need, we developed two plugins within ImageJ for the sole task of automated hemocytometer (or known volume) and migration/invasion cell counting. Likewise, counting membranes from migration/invasion assays with the ImageJ plugin Cell Counter, although accurate, is exceptionally labor intensive, subjective, and infamous for causing wrist pain. When counting large numbers of cell samples, the hemocytometer presents a bottleneck with regards to time. ImageJ has comprehensive particle analysis algorithms which can be used effectively to count various biological particles. The National Institute of Health's ImageJ is a powerful, freely available image processing software suite.
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