title: Development of a single-cell X-ray fluorescence flow cytometer.
abstract: An X-ray fluorescence flow cytometer that can determine the total metal content of single cells has been developed. Capillary action or pressure was used to load cells into hydrophilic or hydrophobic capillaries, respectively. Once loaded, the cells were transported at a fixed vertical velocity past a focused X-ray beam. X-ray fluorescence was then used to determine the mass of metal in each cell. By making single-cell measurements, the population heterogeneity for metals in the µM to mM concentration range on fL sample volumes can be directly measured, a measurement that is difficult using most analytical methods. This approach has been used to determine the metal composition of 936 individual bovine red blood cells (bRBC), 31 individual 3T3 mouse fibroblasts (NIH3T3) and 18 Saccharomyces cerevisiae (yeast) cells with an average measurement frequency of ∼4 cells min(-1). These data show evidence for surprisingly broad metal distributions. Details of the device design, data analysis and opportunities for further sensitivity improvement are described.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Crawford
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Kurecka
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: Tsz Kwan
authors.initials: TK
authors.lastname: Yim
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: Claire
authors.initials: C
authors.lastname: Kozemchak
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: Aniruddha
authors.initials: A
authors.lastname: Deb
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: Lubomír
authors.initials: L
authors.lastname: Dostál
authors.affiliation: X-ray Science Division, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439, USA.
authors.forename: Cheng Jun
authors.initials: CJ
authors.lastname: Sun
authors.affiliation: X-ray Science Division, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439, USA.
authors.forename: Dale L
authors.initials: DL
authors.lastname: Brewe
authors.affiliation: Department of Physics, DePaul University, Chicago, IL 60604, USA.
authors.forename: Raul
authors.initials: R
authors.lastname: Barrea
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Penner-Hahn
mesh_terms: D000818:Animals; D002417:Cattle; D004867:Equipment Design; D005434:Flow Cytometry; D005453:Fluorescence; D051379:Mice; D041681:NIH 3T3 Cells; D011859:Radiography; D014965:X-Rays
keywords: X-ray fluorescence; flow cytometry; homeostasis; metallome; single cell