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An optical counting technique with vertical hydrodynamic focusing for biological cells.
MedLine Citation:
PMID:  20697579     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
A BARRIER IN SCALING LABORATORY PROCESSES INTO AUTOMATED MICROFLUIDIC DEVICES HAS BEEN THE TRANSFER OF LABORATORY BASED ASSAYS: Where engineering meets biological protocol. One basic requirement is to reliably and accurately know the distribution and number of biological cells being dispensed. In this study, a novel optical counting technique to efficiently quantify the number of cells flowing into a microtube is presented. REH, B-lymphoid precursor leukemia, are stained with a fluorescent dye and frames of moving cells are recorded using a charge coupled device (CCD) camera. The basic principle is to calculate the total fluorescence intensity of the image and to divide it by the average intensity of a single cell. This method allows counting the number of cells with an uncertainty +/-5%, which compares favorably to the standard biological methodology, based on the manual Trypan Blue assay, which is destructive to the cells and presents an uncertainty in the order of 20%. The use of a microdevice for vertical hydrodynamic focusing, which can reduce the background noise of out of focus cells by concentrating the cells in a thin layer, has further improved the technique. Computational fluid dynamics (CFD) simulation and confocal laser scanning microscopy images have shown an 82% reduction in the vertical displacement of the cells. For the flow rates imposed during this study, a throughput of 100-200 cellss is achieved.
Authors:
Stefano Chiavaroli; David Newport; Bernie Woulfe
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Publication Detail:
Type:  Journal Article     Date:  2010-06-15
Journal Detail:
Title:  Biomicrofluidics     Volume:  4     ISSN:  1932-1058     ISO Abbreviation:  Biomicrofluidics     Publication Date:  2010  
Date Detail:
Created Date:  2010-08-10     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101293825     Medline TA:  Biomicrofluidics     Country:  United States    
Other Details:
Languages:  eng     Pagination:  -     Citation Subset:  -    
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