Research Papers: General

In situ label-free static cytometry by monitoring spatiotemporal fluctuations of image gray values

[+] Author Affiliations
Ishay Wohl, Naomi Zurgil, Yaron Hakuk, Maria Sobolev, Moti Galmidi, Mordechai Deutsch

Bar Ilan University, The Biophysical Interdisciplinary Schottenstein Center for the Research and Technology of the Cellome, Physics Department, Ramat-Gan 5290002, Israel

J. Biomed. Opt. 20(10), 105013 (Oct 27, 2015). doi:10.1117/1.JBO.20.10.105013
History: Received June 24, 2015; Accepted August 25, 2015
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Abstract.  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.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Ishay Wohl ; Naomi Zurgil ; Yaron Hakuk ; Maria Sobolev ; Moti Galmidi, et al.
"In situ label-free static cytometry by monitoring spatiotemporal fluctuations of image gray values", J. Biomed. Opt. 20(10), 105013 (Oct 27, 2015). ; http://dx.doi.org/10.1117/1.JBO.20.10.105013


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