Hyperspectral image analysis of live cells in various cell cycle stages.
In this study we have explored the use of hyperspectral imaging (HSI) to determine the cell cycle status of live cells in culture. Live cancer cell lines in culture were either synchronized by release from nocodazole or arrested in various cell-cycle phases with serum starvation (G1), aphidicolin (S), or nocodazole (G2/M). The live cells were then stained with the fluorescent DNA binding dyes Heochst 33342 or DCO along with propidium iodide or MTR. Samples were examined using fluorescence microscopy and entire spectral emission profiles were acquired for each sample using a PARISS HSI system. Classified spectra were incorporated into spectral libraries. All spectra acquired from each sample were correlated with library spectra to a user-determined confidence threshold, generating unique spectral signatures for each sample. Examination of these spectral signatures revealed that all cell cycle phases could be objectively differentiated. Ongoing studies employing other viable cell fluorescent dyes, and dyes in combination may provide more robust spectral signatures defining the status and condition of living cells.
Dicker
David T
DT
Laboratory of Molecular Oncology and Cell Cycle Regulation, Department of Medicine (Hematology/Oncology), The Institute for Translational Medicine and Therapeutics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
Lerner
Jeremy M
JM
El-Deiry
Wafik S
WS
D002453:Cell Cycle; D045744:Cell Line, Tumor; D002470:Cell Survival; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation* / Q000379:methods*