Electrode microchamber for noninvasive perturbation of mammalian cells with nanosecond pulsed electric fields.
Nanosecond pulsed electric fields can pass through the external membrane of biological cells and disturb fast-responding intracellular structures and processes. To enable real-time imaging and investigation of these phenomena, a microchamber with integral electrodes and optical path for observing individual cells exposed to ultrashort electric pulses was designed and fabricated utilizing photolithographic and microelectronic methods. SU-8 photoresist was patterned to form straight sidewalls from 10 to 30 microm in height, with gold film deposited on the top and sidewalls for conductive, nonreactive electrodes and a uniform electric field. Channel dimensions (10-30 microm x 100 microm x 12 000 microm) are suitable for observations of mammalian cells during nanosecond, megavolt-per-meter pulsed electric field exposure. Experimental studies utilizing the electrode microchamber include live-cell imaging of nanoelectropulse-induced intracellular calcium bursts and membrane phospholipid translocation.
Sun
Yinghua
Y
Materials Science Department, University of Southern California, Los Angeles, CA 90089, USA. yinghuas@usc.edu
Vernier
P Thomas
PT
Behrend
Matthew
M
Marcu
Laura
L
Gundersen
Martin A
MA
D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D004558:Electric Stimulation / Q000295:instrumentation* / Q000379:methods; D004574:Electromagnetic Fields; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D019169:Jurkat Cells; D008839:Microelectrodes; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods
D002118:Calcium