Magnetic flow sorting using a model system of human lymphocytes and a colloidal magnetic label.
Cells of identical physical properties that differ in the expression of surface proteins can be sorted conveniently using immunospecific stains conjugated to fluorescent, or magnetic, labels. Immunomagnetic cell sorting using commercial batch sorters offers advantages of high sorting capacity, high viability of sorted fractions, and high depletion rates; its disadvantages are low enrichment rate and batch processing. The authors developed and tested a continuous, flow-through magnetic cell sorter for small volume, experimental cell enrichment. Freshly isolated human peripheral lymphocytes were labeled using an immunofluoromagnetic sandwich consisting of mouse anti human CD8 monoclonal antibody-fluorescein conjugate and rat anti mouse polyclonal antibody-colloidal iron-dextran conjugate. A total of 2-3 min lymphocytes were sorted per hour using a saturation magnetic field of 1.334 T and a five channel sorter. The fluorescent cells were distributed among the channels in relation to their fluorescence intensity and magnetic susceptibility. The purity (68-85%) and enrichment rates (16-34x) were comparable to those of commercial batch magnetic separators; sorting capacity and recovery of the enriched fractions (up to 32%) were limited by the small scale of the sorter. Future direction is focused on increasing the resolution, recovery, and sorting capacity of the enriched fractions, and testing the sorter on other cell systems.
Zborowski
M
M
Department of Biomedical Engineering, Cleveland Clinic Foundation, OH 44195-5254, USA.
Moore
L R
LR
Reddy
S
S
Chen
G H
GH
Sun
L
L
Chalmers
J J
JJ
D000818:Animals; D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D003102:Colloids; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation / Q000379:methods*; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D051381:Rats
D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; D003102:Colloids