On-chip cell lysis by local hydroxide generation.
We present a novel method for on-chip cell lysis based on local hydroxide electro-generation. Hydroxide ions porate the cell membrane, leading to cell lysis. After lysis occurs, hydrogen ions, also generated on chip, react with excess hydroxide, creating a neutral pH lysate and eliminating the need for a wash step. Three different cell types are shown to be effectively lysed by this method: red blood cells, HeLa (human tumor line) and Chinese Hamster Ovary (CHO) cell lines. The release of cytoplasmic molecules from HeLa and CHO cells is demonstrated by monitoring the escape of a membrane impermeant dye from the cytoplasm. In the vicinity of the cathode, the hydroxide concentration is predicted by finite element simulations and shown to fit the lysis rates at different distances from the generating cathode. For flow-through experiments, a second device integrating a mechanical filter with hydroxide generation is fabricated and tested. The purpose of the filter is to trap whole cells and only allow lysate to pass through. The flow rate dependence of hydroxide concentration at the lysis filter is modeled and lysis efficiency is experimentally determined to be proportional to the hydroxide concentration for flow rates from 15 to 30 microl min(-1).
Di Carlo
Dino
D
Department of Bioengineering, University of California at Berkeley, Berkeley, CA, USA.
Ionescu-Zanetti
Cristian
C
Zhang
Yolanda
Y
Hung
Paul
P
Lee
Luke P
LP
D000818:Animals; D002460:Cell Line; D002477:Cells / Q000737:chemistry* / Q000378:metabolism; D006224:Cricetinae; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006878:Hydroxides / Q000032:analysis* / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D012756:Sheep
D006878:Hydroxides