Control of the Release of Freely Diffusing Molecules in Single-Cell Electroporation

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Control of the Release of Freely Diffusing Molecules in Single-Cell Electroporation

Agarwal, A. (författare): University of Pittsburgh

Wang, M. Y. (författare): University of Pittsburgh

Olofsson, Jessica, 1975 (författare): Chalmers tekniska högskola,Chalmers University of Technology

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Orwar, Owe, 1964 (författare): Chalmers tekniska högskola,Chalmers University of Technology

Weber, S. G. (författare): University of Pittsburgh

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(creator_code:org_t)

2009-09-04
2009
Engelska.
Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 81:19, s. 8001-8008

Relaterad länk:: https://europepmc.or...; visa fler...; https://doi.org/10.1...; https://research.cha...; visa färre...

Tidskriftsartikel (refereegranskat)

Abstract Ämnesord

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Single-cell electroporation using an electrolyte-filled capillary is an emerging technique for transient pore formation in adherent cells. Because adherent cells do not have a simple and consistent shape and because the electric field emanating from the tip of the capillary is inhomogeneous, the Schwan equation based on spherical cells in homogeneous electrical fields does not apply. We sought to determine experimental and cell parameters that influence the outcome of a single-cell electroporation experiment. A549 cells were exposed to the thiol-reactive dye Thioglo-1, leading to green fluorescence from intracellular thiol adducts. Electroporation causes a decrease with time of the intracellular fluorescence intensity of Thioglo-1-loaded cells from diffusive loss of thiol adducts. The transient curves thus obtained are well-described by a simple model originally developed by Puc et al. We find that the final fluorescence following electroporation is related to the capillary tip-to-cell distance and cell size (specifically, 2(A/pi)(1/2) where A is the area of the cell's image in pixels. This quantity is the diameter if the image is a circle). In separate experiments, the relationship obtained can be used to control the final fluorescence following electroporation by adjusting the tip-to-cell distance based on cell size. The relationship was applied successfully to A549 as Well as DU 145 and PC-3 cells. Finally, F-tests show that the variability in the final fluorescence (following electroporation) is decreased when the tip-to-cell distance is controlled according to the derived relationship in comparison to experiments in which the tip-cell distance is a constant irrespective of cell size.

Ämnesord

NATURVETENSKAP -- Kemi -- Fysikalisk kemi (hsv//swe)
NATURAL SCIENCES -- Chemical Sciences -- Physical Chemistry (hsv//eng)

Nyckelord

membrane
gene-transfer
drug-delivery
induced dna transfection
mammalian-cells
electropermeabilization
electric-field
electrolyte-filled capillary
diameter capillaries
human erythrocytes
chemical-analysis

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Av författaren/redakt...: Agarwal, A.; Wang, M. Y.; Olofsson, Jessic ...; Orwar, Owe, 1964; Weber, S. G.

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NATURVETENSKAP: NATURVETENSKAP; och Kemi; och Fysikalisk kemi

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Av lärosätet: Chalmers tekniska högskola

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