Microfluidic devices for the high-throughput chemical analysis of cells.
A microfluidic device is reported that integrated cell handling, rapid cell lysis, and electrophoretic separation and detection of fluorescent cytosolic dyes. The device function was demonstrated using Jurkat cells that were loaded with the fluorogenic dyes - carboxyfluorescein diacetate, Oregon green carboxylic acid diacetate, or Calcein AM. The loaded cells were hydrodynamically transported from the cell-containing reservoir to a region on the microfluidic device where they were focused and then rapidly lysed using an electric field. Complete lysis was accomplished in <33 ms. The hydrolyzed, fluorescent dyes in the cell lysate were automatically injected into a separation channel on the device and detected 3 mm downstream of the injection point. The total separation time was approximately 2.2 s with absolute migration time reproducibilities of <1% and efficiencies ranging from 2300 to 4000 theoretical plates. Results from 139 cells are reported. A small fraction of these cells, approximately 9%, were found to enzymatically hydrolyze the loaded dyes in a manner significantly different from the majority of the cells. Cell analysis rates of 7-12 cells/min were demonstrated and are >100 times faster than those reported using standard bench-scale capillary electrophoresis.
Oak Ridge National Laboratory, P.O. Box 2008, Oak Ridge, Tennessee 37831-6142, USA.
Maxine A
MA
McClain
Christopher T
CT
Culbertson
Stephen C
SC
Jacobson
Nancy L
NL
Allbritton
Christopher E
CE
Sims
J Michael
JM
Ramsey
D000465:Algorithms; D002264:Carboxylic Acids / Q000032:analysis / Q000378:metabolism; D002448:Cell Adhesion; D002452:Cell Count; D002458:Cell Fractionation / Q000379:methods; D002477:Cells / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D004572:Electrolysis; D004586:Electrophoresis / Q000379:methods; D004867:Equipment Design; D005452:Fluoresceins / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis*; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D018715:Microscopy, Video; D012212:Rheology; D013050:Spectrometry, Fluorescence
D002264:Carboxylic Acids; D005452:Fluoresceins; D005456:Fluorescent Dyes; C436864:Oregon Green 488 carboxylic acid; C027780:carboxyfluoresceindiacetate; C085925:calcein AM