title: The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay.
abstract: Cytometric studies utilizing flow cytometry or multi-well culture plate fluorometry are often limited by a deficit in temporal resolution and a lack of single cell consideration. Unfortunately, many cellular processes, including signaling, motility, and molecular transport, occur transiently over relatively short periods of time and at different magnitudes between cells. Here we demonstrate the multitrap nanophysiometer (MTNP), a low-volume microfluidic platform housing an array of cell traps, as an effective tool that can be used to study individual unattached cells over time with precise control over the intercellular microenvironment. We show how the MTNP platform can be used for hematologic cancer cell characterization by measuring single T cell levels of CRAC channel modulation, non-translational motility, and ABC-transporter inhibition via a calcein-AM efflux assay. The transporter data indicate that Jurkat T cells exposed to indomethacin continue to accumulate fluorescent calcein for over 60 minutes after calcein-AM is removed from the extracellular space.
authors.affiliation: 1] Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA [2] University of New Mexico School of Medicine, Albuquerque, NM, 87131, USA.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Byrd
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Loi T
authors.initials: LT
authors.lastname: Hoang
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Eric G
authors.initials: EG
authors.lastname: Kim
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Pfister
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Erik M
authors.initials: EM
authors.lastname: Werner
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Arndt
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Chamberlain
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Jacob J
authors.initials: JJ
authors.lastname: Hughey
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Bao A
authors.initials: BA
authors.lastname: Nguyen
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Erik J
authors.initials: EJ
authors.lastname: Schneibel
authors.affiliation: Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Laura L
authors.initials: LL
authors.lastname: Wertz
authors.affiliation: Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Whitfield
authors.affiliation: 1] Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA [2] Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, 37235, USA [3] Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37235, USA [4] Department of Physics and Astronomy, Vanderbilt University, Nashville, TN, 37235, USA [5] Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Wikswo
authors.affiliation: 1] Searle Systems Biology and Bioengineering Undergraduate Research Experience, Vanderbilt University, Nashville, TN, 37235, USA [2] Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, 37235, USA [3] Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37235, USA.
authors.forename: Kevin T
authors.initials: KT
authors.lastname: Seale
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002469:Cell Separation / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005434:Flow Cytometry / Q000295:instrumentation*; D005452:Fluoresceins / Q000032:analysis; D006801:Humans; D019169:Jurkat Cells; D015458:Leukemia, T-Cell / Q000378:metabolism* / Q000473:pathology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D036103:Nanotechnology / Q000295:instrumentation; D052898:Optical Tweezers; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D046888:Tissue Array Analysis / Q000295:instrumentation*
chemical_list: D002135:Calcium-Binding Proteins; D005452:Fluoresceins; C085925:calcein AM