The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay

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 rel...

Full description

Saved in:

Bibliographic Details
Published in:	Scientific reports Vol. 4; no. 1; p. 5117
Main Authors:	Byrd IV, Thomas F., Hoang, Loi T., Kim, Eric G., Pfister, Matthew E., Werner, Erik M., Arndt, Stephen E., Chamberlain, Jeffrey W., Hughey, Jacob J., Nguyen, Bao A., Schneibel, Erik J., Wertz, Laura L., Whitfield, Jonathan S., Wikswo, John P., Seale, Kevin T.
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 30-05-2014 Nature Publishing Group
Subjects:	13/62 14/34 631/1647/664/1257 631/1647/664/1364 Animals Blood cancer Calcein Calcium channels Calcium-Binding Proteins - metabolism Cancer Cell culture Cell Separation - instrumentation Equipment Design Equipment Failure Analysis Flow cytometry Flow Cytometry - instrumentation Fluoresceins - analysis Fluorometry Humanities and Social Sciences Humans Indomethacin Jurkat Cells Leukemia, T-Cell - metabolism Leukemia, T-Cell - pathology Lymphocytes T Microfluidic Analytical Techniques - instrumentation Microfluidics Motility multidisciplinary Nanotechnology - instrumentation Optical Tweezers Reproducibility of Results Science Sensitivity and Specificity Tissue Array Analysis - instrumentation
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	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.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-4 ObjectType-Undefined-1 content type line 23 ObjectType-Undefined-3
ISSN:	2045-2322 2045-2322
DOI:	10.1038/srep05117