FLOW CYTOMETRY IN IMMUNOLOGY AND HEMATOLOGY 

Minerva Biotecnologica 1999 June;11(2):93-100

Copyright © 1999 EDIZIONI MINERVA MEDICA

language: English

Multiparametric analysis in flow cytometry

Ortolani C.

Flow Cytometry Unit, Laboratory of Clinical Pathology, Venice General Hospital, Venice, Italy

Flow cytom­e­try has ­been deep­ly mod­i­fied by a con­tin­u­ous ­series of advanc­es regard­ing the ­most ­diverse ­fields of mod­ern tech­nol­o­gy. The ­most impor­tant improve­ment con­sists in its capac­ity to inves­ti­gate sev­er­al dif­fer­ent param­e­ters in the ­same ana­lyt­i­cal run, and ­then repro­cess the ­data in a mul­ti­var­i­ate anal­y­sis. This impor­tant func­tion has ­been ­allowed not ­only by the avail­abil­ity of mul­ti­chan­nel ­flow cytom­e­ters, but ­also by the syn­the­sis of flu­o­res­cent ­dyes ­with con­ven­ient­ly ­matched spec­tra. Although ­very pop­u­lar, nev­er­the­less a mul­ti­par­a­met­ric ­approach is not a triv­i­al ­task, inas­much it ­relies on the ­firm con­trol of ­each ana­lyt­i­cal var­i­able and pre­sup­pos­es the fix­ing of the arte­facts ­which can inter­fere ­with the meas­ures. In ­this arti­cle ­some piv­ot­al argu­ments are dis­cussed, i.e. the ­choice of flu­o­roch­romes, the con­trol of arte­facts, and the anal­y­sis of ­data. Special empha­sis has ­been put on the ­more ­tricky sit­u­a­tions, ­like spec­tral and ster­i­cal incom­pat­ibil­ity ­between ­dyes, and the ­spill-­over ­between dif­fer­ent chan­nels.