Special Section on Quantitative Phase Imaging in Biomedicine

Differentiating neutrophils using the optical coulter counter

[+] Author Affiliations
Ethan Schonbrun, Giuseppe Di Caprio

Harvard University, Rowland Institute, 100 Edwin H. Land Boulevard, Cambridge, Massachusetts 02142, United States

J. Biomed. Opt. 20(11), 111205 (Jul 17, 2015). doi:10.1117/1.JBO.20.11.111205
History: Received March 15, 2015; Accepted June 15, 2015
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Abstract.  We present an optofluidic measurement system that quantifies cell volume, dry mass, and nuclear morphology of neutrophils in high-throughput. While current clinical hematology analyzers can differentiate neutrophils from a blood sample, they do not give other quantitative information beyond their count. In order to better understand the distribution of neutrophil phenotypes in a blood sample, we perform two distinct multivariate measurements. In both measurements, white blood cells are driven through a microfluidic channel and imaged while in flow onto a color camera using a single exposure. In the first measurement, we quantify cell volume, scattering strength, and cell dry mass by combining quantitative phase imaging with dye exclusion cell volumetric imaging. In the second measurement, we quantify cell volume and nuclear morphology using a nucleic acid fluorescent stain. In this way, we can correlate cell volume to other cellular characteristics, which would not be possible using an electrical coulter counter. Unlike phase imaging or cell scattering analysis, the optical coulter counter is capable of quantifying cell volume virtually independent of the cell’s refractive index and unlike optical tomography, measurements are possible on quickly flowing cells, enabling high-throughput.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Ethan Schonbrun and Giuseppe Di Caprio
"Differentiating neutrophils using the optical coulter counter", J. Biomed. Opt. 20(11), 111205 (Jul 17, 2015). ; http://dx.doi.org/10.1117/1.JBO.20.11.111205


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