title: Field-flow fractionation of cells with chemiluminescence detection.
abstract: Field-flow fractionation is a separation technique characterized by a retention mechanism which makes it suitable for sorting cells over a short analysis time, with low sample carry-over and preserving cell viability. Thanks to its high sensitivity, chemiluminescence detection is suitable for the quantification of just a few cells expressing chemiluminescence or bioluminescence. In this work, different formats for coupling gravitational field-flow fractionation and chemiluminescence detection are explored to achieve ultra-sensitive cell detection in the framework of cell sorting. The study is carried out using human red blood cells as model sample. The best performance is obtained with the on-line coupling format, performed in post-column flow-injection mode. Red cells are isolated from diluted whole human blood in just a few minutes and detected using the liquid phase chemiluminescent reaction of luminol catalysed by the red blood cell heme. The limit of detection is a few hundred injected cells. This is lower than the limit of detection usually achieved by means of conventional colorimetric/turbidimetric methods, and it corresponds to a red blood cell concentration in the injected sample of five orders of magnitude lower than in whole blood.
authors.affiliation: Department of Chemistry G. Ciamician, University of Bologna, Via Selmi 2, I-40126 Bologna, Italy. dora.melucci@unibo.it
authors.forename: Dora
authors.initials: D
authors.lastname: Melucci
authors.forename: Barbara
authors.initials: B
authors.lastname: Roda
authors.forename: Andrea
authors.initials: A
authors.lastname: Zattoni
authors.forename: Sonia
authors.initials: S
authors.lastname: Casolari
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Reschiglian
authors.forename: Aldo
authors.initials: A
authors.lastname: Roda
mesh_terms: D004912:Erythrocytes / Q000166:cytology*; D031521:Fractionation, Field Flow; D006418:Heme / Q000737:chemistry; D006801:Humans; D049449:Luminescence; D008165:Luminol / Q000737:chemistry; D012680:Sensitivity and Specificity
chemical_list: D006418:Heme; D008165:Luminol