High-content Screening Method for Detecting Cell Proliferation Cycle
Molecular Screening: siRNA libraries aiming at major human diseases
Cell Model: human cervical carcinoma cell Hela
Detection Instrument: high-content screening system
Detection Index: cell proliferation, cell cycle
Cell culture; cell transfection; cell staining; fluorescence scanning
Put the scanned data into different groups according to the cell size (width, depth) and fluorescence intensity (total intensity), and obtain the distribution map and data for different cells. From the figure below, it shows that comparing with NC control group, the total cell counts are remarkably decreased after adding siRNA for gene 2, which indicates that the siRNA for gene2 inhibits Hela cell proliferation.
2) Analyze the influences on total cell counts after adding siRNA from the statistical analysis of data
Perform the statistical analysis after analyzing cells. It shows that siRNAs aiming at gene1, 8, 9, 10 obviously promote the cell proliferation, while siRNAs for gene 2, 3, 4, 5, 6, 7 significantly inhibit cell proliferation and siRNA for gene11 has no obvious influence on cell proliferation when comparing with NC control group.
3) Analyze the influences on cell cycles after adding siRNA from the statistical analysis of data
Perform the statistical analysis after analyzing cells. It shows that siRNAs aiming at gene1, 8, 9, 10, 11 obviously increase cell counts at the G1 phase, while siRNAs for gene 2, 3, 4, 5, 6, 7 significantly increase cells at the G2/M phase when comparing with NC control group. From the cell distribution frequency diagram of gene1 and gene2 with different fluorescence intensity, we can also see that the cells treated by gene1 siRNA are remarkably more than gene 2 at the G1 phase, while cells at the G2/M phase are clearly less than gene 2 siRNA.
4) 3D cell image
During the experiment, 3D cell image is also provided, which can be used to study the cell location in the cell signaling transduction.
5) Cell heatmap after the siRNA treatment
Molecular drug with obvious differences will be easily found by using Heatmap. Below is the shown heatmap of total cell counts and G1 cell counts.