Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers.
Phenotypically pure subpopulations of lymphocytes can provide valuable insights into the immune response to injury and disease. The isolation of these subpopulations presents unique challenges, particularly when preprocessing incubation to attach fluorescent or antibody tags is to be minimized. This paper examines the separation of T and B lymphocytes from mixtures using microfluidic chambers coated with antibodies, focusing on flow conditions and surface chemistry. The adhesion of both cell types decreases as shear stress increases irrespective of the surface chemistry. The incorporation of poly(ethylene glycol) chains along with the antibodies on the chamber surface is shown to significantly improve the reproducibility of cell adhesion and is thus an important part of the overall system design. Furthermore, this technique is shown to be an effective way of isolating highly pure subpopulations of lymphocytes from model mixtures, even when the target cell concentration is low.
Murthy
Shashi K
SK
Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
Sin
Aaron
A
Tompkins
Ronald G
RG
Toner
Mehmet
M
D000906:Antibodies / Q000276:immunology; D001360:Avidin / Q000737:chemistry; D018956:CD5 Antigens / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D008214:Lymphocytes / Q000166:cytology*; D044085:Microfluidics / Q000295:instrumentation*; D018625:Microscopy, Atomic Force; D011092:Polyethylene Glycols / Q000737:chemistry; D012212:Rheology
D000906:Antibodies; D018956:CD5 Antigens; D001360:Avidin; D011092:Polyethylene Glycols