Spatial control of cellular measurements with the laser micropipet

Analytical Chemistry
H LiN L Allbritton

Abstract

Continued progress in understanding cellular physiology requires new strategies for biochemical measurements in solitary cells, multiple cells, and subcompartments of cells. Large spatial gradients in the concentrations of molecules and presumably the activities of enzymes can occur in cells. Consequently, there is a critical need for measurement techniques for mammalian cells with control over the numbers or regions of cells interrogated. In the present work, we developed a strategy to rapidly load the cytoplasmic contents of either multiple cells or a subregion of a single cell into a capillary. A single, focused pulse from a laser created a mechanical shock wave which disrupted a group of cells or a portion of a cell in the path of the shock wave. Simultaneously, the cytoplasm was loaded into a capillary for electrophoretic separation. The size of the region of cellular disruption (and therefore the volume of cytoplasm collected) was controlled by the amount of energy in the laser pulse. Higher energies could be used to sample groups of cells while much lower energies could be utilized to selectively sample the tip of a neuronal process. The feasibility of performing measurements on subcellular compartments was also demonstr...Continue Reading

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Citations

Feb 24, 2005·Analytical and Bioanalytical Chemistry·Paula R Powell, Andrew G Ewing
Aug 29, 2006·Analytical and Bioanalytical Chemistry·Ryan D JohnsonEdgar A Arriaga
Oct 29, 2003·Current Opinion in Chemical Biology·Norman J Dovichi, Shen Hu
Aug 22, 2003·Journal of Chromatography. a·Yan WangChristopher E Sims
Feb 5, 2003·Current Opinion in Biotechnology·Christopher E Sims, Nancy L Allbritton
Nov 6, 2007·Analytical Chemistry·Paul J MarcNancy L Allbritton
Apr 25, 2008·Lab on a Chip·Paul J MarcNancy L Allbritton
Apr 23, 2008·Journal of the Royal Society, Interface·Robert B Brown, Julie Audet
Jun 28, 2008·Journal of the Royal Society, Interface·Hsuan-Hong LaiNancy L Allbritton
Jul 19, 2008·Annual Review of Analytical Chemistry·Laura M BorlandNancy L Allbritton
Jul 18, 2014·Proceedings of the National Academy of Sciences of the United States of America·Zhen YangZhifeng Ren
Feb 13, 2016·Biomicrofluidics·Guillaume LajoinieMichel Versluis
Feb 27, 2010·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Robert B BrownJulie Audet
Oct 24, 2003·Analytical Chemistry·Futian HanNancy L Allbritton
Nov 1, 2003·Analytical Chemistry·Maxine A McClainJ Michael Ramsey
May 30, 2003·Analytical and Bioanalytical Chemistry·Lori A Woods, Andrew G Ewing
Mar 29, 2007·Lab on a Chip·Christopher E Sims, Nancy L Allbritton
Jun 23, 2007·Biomedical Chromatography : BMC·Suresh C V BabuYoung Sook Yoo
Jun 20, 2007·Electrophoresis·Yun ChenEdgar A Arriaga
Apr 4, 2008·Electrophoresis·Hui YanHongkai Wu
Sep 4, 2015·Journal of Biosciences·Guido A Drexler, Miguel J Ruiz-Gomez
May 21, 2013·Analytical Chemistry·Ryan M PhillipsNancy L Allbritton
Sep 15, 2012·Analytical Chemistry·Yihong ZhanChang Lu

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