Document Detail

Continuous flow microfluidic device for rapid erythrocyte lysis.
MedLine Citation:
PMID:  15516115     Owner:  NLM     Status:  MEDLINE    
Leukocyte isolation from whole blood to study inflammation requires the removal of contaminating erythrocytes. Leukocytes, however, are sensitive to prolonged exposure to hyper/hypoosmotic solutions, temperature changes, mechanical manipulation, and gradient centrifugation. Even though care is taken to minimize leukocyte activation and cell loss during erythrocyte lysis, it is often not possible to completely avoid it. Most procedures for removal of contaminating erythrocytes from leukocyte preparations are designed for bulk processing of blood, where the sample is manipulated for longer periods of time than necessary at the single-cell level. Ammonium chloride-mediated lysis is the most commonly used method to obtain enriched leukocyte populations but has been shown to cause some activation and selective loss of certain cell types. The leukocyte yield and subsequent activation status of residual leukocytes after NH(4)Cl-mediated lysis have been shown to depend on the time of exposure to the lysis buffer. We have developed a microfluidic lysis device that deals with erythrocyte removal at nearly the single-cell level. We can achieve complete lysis of erythrocytes and approximately 100% recovery of leukocytes where the cells are exposed to an isotonic lysis buffer for less than 40 s, after which the leukocytes are immediately returned to physiological conditions. Theoretically, this process can be made massively parallel to process several milliliterss of whole blood to obtain a pure leukocyte population in less than 15 min.
Palaniappan Sethu; Melis Anahtar; Lyle L Moldawer; Ronald G Tompkins; Mehmet Toner
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical chemistry     Volume:  76     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2004 Nov 
Date Detail:
Created Date:  2004-11-01     Completed Date:  2006-06-21     Revised Date:  2013-12-03    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  6247-53     Citation Subset:  IM    
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MeSH Terms
Microfluidics / instrumentation*
Rats, Inbred Lew
Grant Support

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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