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Quantification of Live Cell Imaging

Live cell imaging is a useful tool to monitor cellular activities in living systems. It is often necessary in cancer research or experimental research to quantify the dividing capabilities of cells or the cell proliferation level when investigating manipulations of the cells or their environment. Manual quantification of fluorescence microscopic image is difficult because human is neither sensitive to fine differences in color intensity nor effective to count and average fluorescence level among cells. However, auto-quantification is not a straightforward problem to solve. As the sampling location of the microscopy changes, the amount of cells in individual microscopic images varies, which makes simple measurement methods such as the sum of stain intensity values or the total number of positive stain within each image inapplicable. Thus, automated quantification with robust cell segmentation techniques is required. Here, an unsupervised entropy based cell segmentation is developed and a robust automated quantification method of live cell imaging is built to measure the cell replication level, providing a robust quantitative analysis system in fluorescent live cell imaging. The experimental results in application to monitor cellular replication activities show that the quantitative score is promising to represent the cell replication level, and scores for images from different cell replication groups are demonstrated to be statistically significantly different using ANOVA and LSD tests. In addition, the technique is fast and takes less than 0.5 second for high resolution microscopic images.


This software is released with the manuscript (Fast Automatic Quantitative Cell Replication with Fluorescent Live Cell Imaging). You are free to download and use it for research purposes. The software is developed in java (with jdk1.6.0_22 installed) and based on ImageJ. Therefore, please install jdk before use it.

A zip file is available here. You can decompress the file and double click RunMe.bat. Then, you can activate the program.

Test Images

Links to some test images are in the right column.

Simple steps to use the software

  • install jdk
  • download the zip file and the images
  • double click RunMe.bat
  • click file=>open => open the test images
  • click plugin=>filter=>Quantify Cell Color
  • The quantification output is displayed in a log window.