By Topic

Intra-microfluidic pinocytic loading of human T cells

Sign In

Cookies must be enabled to login.After enabling cookies , please use refresh or reload or ctrl+f5 on the browser for the login options.

Formats Non-Member Member
$31 $13
Learn how you can qualify for the best price for this item!
Become an IEEE Member or Subscribe to
IEEE Xplore for exclusive pricing!
close button

puzzle piece

IEEE membership options for an individual and IEEE Xplore subscriptions for an organization offer the most affordable access to essential journal articles, conference papers, standards, eBooks, and eLearning courses.

Learn more about:

IEEE membership

IEEE Xplore subscriptions

3 Author(s)
Hughey, J.J. ; Vanderbilt Univ., Nashville ; Wikswo, John P. ; Seale, K.T.

Pinocytic loading (PL) exposes cells sequentially to hypertonic (0.5 M sucrose and 10% PEG 1000) and hypotonic (60% v/v media in water) solutions to transfer cell-impermeant macromolecules into the cytosol. Here we leverage the advantages of microfluidics to achieve high-efficiency pinocytic loading of Jurkat T cells with fluorescein dextran. To our knowledge, we report the first observation of the time course of PL in single cells. In contrast to conventional PL, cells in the microfluidic device can be rapidly cycled through the various solutions. While the conventional protocol provides only 40-60% efficiency, we have achieved over 90% loading efficiency in the microfluidic device. Mean cell fluorescence from conventional PL, is approximately half that of microfluidic PL with three 10-minute hypertonic cycles (49.7plusmn21.8 and 21.7plusmn17.8 a.u. respectively, P=4ldr10-17). While the mean and median cell fluorescence increases with each cycle in the microfluidic device, fluorescence begins to plateau after three 10-minute hypertonic cycles; subsequent cycles see dramatic increases in cell death from 10% to over 30%. Hypotonic media does not significantly affect loading efficiency for intra-inicrofluidic PL, but causes slightly higher cell death. We believe that intra-microfluidic PL of living cells is a valuable tool for BioMEMS-based systems biology.

Published in:

Life Science Systems and Applications Workshop, 2007. LISA 2007. IEEE/NIH

Date of Conference:

8-9 Nov. 2007