Fully-automated image processing software to analyze calcium traces in populations of single cells.
Advances in fluorescence live cell imaging over the last decade have revolutionized cell biology by providing access to single-cell information in space and time. One current limitation of live-cell imaging is the lack of automated procedures to analyze single-cell data in large cell populations. Most commercially available image processing softwares do not have built-in image segmentation tools that can automatically and accurately extract single-cell data in a time series. Consequently, individual cells are usually identified manually, a process which is time consuming and inherently low-throughput. We have developed a MATLAB-based image segmentation algorithm that reliably detects individual cells in dense populations and measures their fluorescence intensity over time. To demonstrate the value of this algorithm, we measured store-operated calcium entry (SOCE) in hundreds of individual cells. Rapid access to single-cell calcium signals in large populations allowed us to precisely determine the relationship between SOCE activity and STIM1 levels, a key component of SOCE. Our image processing tool can in principle be applied to a wide range of live-cell imaging modalities and cell-based drug screening platforms.
Wong
Loo Chin
LC
DUKE-NUS Graduate Medical School, Program in Neuroscience and Neurobehavioral Disorders, 8 College Road, Singapore 169857, Singapore.
Lu
Bo
B
Tan
Kia Wee
KW
Fivaz
Marc
M
D000465:Algorithms; D000818:Animals; D002118:Calcium / Q000032:analysis*; D020013:Calcium Signaling / Q000502:physiology; D045744:Cell Line, Tumor; D002999:Clone Cells; D001330:Electronic Data Processing / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D059010:Single-Cell Analysis / Q000379:methods*; D012984:Software; D013223:Statistics as Topic; D000071737:Stromal Interaction Molecule 1; D014162:Transfection
D008562:Membrane Glycoproteins; D034741:RNA, Small Interfering; C518857:Stim1 protein, rat; D000071737:Stromal Interaction Molecule 1; D002118:Calcium