Abstract
Flow cytometry is a powerful tool for cell counting and biomarker detection in biotechnology and medicine especially with regards to blood analysis. Standard flow cytometers perform cell type classification both by estimating size and granularity of cells using forward- and side-scattered light signals and through the collection of emission spectra of fluorescently-labeled cells. However, cell surface labeling as a means of marking cells is often undesirable as many reagents negatively impact cellular viability or provide activating/inhibitory signals, which can alter the behavior of the desired cellular subtypes for downstream applications or analysis. To eliminate the need for labeling, we introduce a label-free imaging-based flow cytometer that measures size and cell protein concentration simultaneously either as a stand-alone instrument or as an add-on to conventional flow cytometers. Cell protein concentration adds a parameter to cell classification, which improves the specificity and sensitivity of flow cytometers without the requirement of cell labeling. This system uses coherent dispersive Fourier transform to perform phase imaging at flow speeds as high as a few meters per second.
Highlights
Cell protein content measurement can be used in many biomedical applications such as blood doping detection [1], infection monitoring [2], drug development and screening [3], studies of necrosis and apoptosis [4, 5], cell cycle progression and differentiation [6,7,8], and in cancer diagnostics [9,10,11]
Flow cytometry is a powerful tool for cell counting and biomarker detection in biotechnology and medicine especially with regards to blood analysis
Cell surface labeling as a means of marking cells is often undesirable as many reagents negatively impact cellular viability or provide activating/inhibitory signals, which can alter the behavior of the desired cellular subtypes for downstream applications or analysis
Summary
Cell protein content measurement can be used in many biomedical applications such as blood doping detection [1], infection monitoring [2], drug development and screening [3], studies of necrosis and apoptosis [4, 5], cell cycle progression and differentiation [6,7,8], and in cancer diagnostics [9,10,11]. Current methods for cell protein concentration measurement include electrical methods based on dielectrophoresis [12], mechanical methods based on microchannel cantilevers [1], and optical methods based on scattering patterns [13], emission spectra of external cavity lasers [14], and holographic and phase microscopy [15,16,17,18]. These methods are either inherently too slow for high-speed flow cytometry applications or require feedback mechanisms [19] to provide necessary precision. The simultaneous measurement of refractive index and size of cells would be predicted to provide two independent parameters for cell classification
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