In situ label-free static cytometry by monitoring spatiotemporal fluctuations of image gray values.
Spatiotemporal fluctuation of homogeneity and randomness of gray values within an image was explored and utilized as a label-free means for cell examination. This was done by utilizing a user-friendly combination of simple bright field microscope and Cytocapture dish, wherein cells are individually held, each within a picoliter optical chamber, forming an array of cells to be repeatedly measured over time and biomanipulated in situ at single-cell resolution. First, the measured gray level information entropy (GLIE) was used and, based on the fact that living cells are not in a state of thermodynamic equilibrium but rather in a metastable state, two fluctuation-sensitive measures were proposed and examined: ASDE—the spatial average of temporal standard deviation (SD) of GLIE, and AA—the average time autocorrelation of GLIE. System performance was validated on cell-free solutions. This was followed by examining the performance of the measures AGLIE, ASDE, and AA to distinguish among individual live-still, dead and live cells from various cell lines, as well as between cells which were and were not induced to differentiate. Results, which were obtained on four types of cells, indicate advantages of the proposed measures which are believed to be significant additions to the microscope-based probe-free toolbox.
Ishay
I
Wohl
Naomi
N
Zurgil
Yaron
Y
Hakuk
Maria
M
Sobolev
Moti
M
Galmidi
Mordechai
M
Deutsch
D002469:Cell Separation / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D005434:Flow Cytometry / Q000295:instrumentation*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D062211:Spatio-Temporal Analysis; D013194:Staining and Labeling; D020298:U937 Cells