Microfluidic selection and retention of a single cardiac myocyte, on-chip dye loading, cell contraction by chemical stimulation, and quantitative fluorescent analysis of intracellular calcium

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Detail(s)

Original languageEnglish
Pages (from-to)4315-4322
Journal / PublicationAnalytical Chemistry
Volume77
Issue number14
Publication statusPublished - 15 Jul 2005
Externally publishedYes

Abstract

A microfluidic method to study the contraction of a single cardiac myocyte (heart muscle cell) has been developed. This method integrates various single-cell operations as well as on-chip dye loading, and quantitative analysis of intracellular calcium concentration, [Ca2+]i. After the channel enlargement by on-chip etching to accommodate large-sized cardiac myoytes, a single cell is selected and retained at a V-shaped cell retention structure within the microchip. Owing to the fragile property of the cardiac myocytes that could easily be damaged by centrifugation, the calcium-sensitive fluorescent dye was loaded in the cell by on-chip dye loading. This on-chip method minimized the damage to the cells from the use of a centrifuge in the conventional method and provided a way of cellular analysis of fragile cells. Subsequently, quantitative analysis of [Ca2+]i of a single cardiac myocyte by fluorescence measurement was achieved for the first time in a microfluidic chip, thanks to the intracellular calcium stimulant of ionomycin. The resting [Ca2+]i of the cardiomyocyte determined was consistent with the literature value. From the spontaneous contraction study, it was found that fluorescence intensity cannot represent the [Ca 2+]i variation accurately, which implied the importance of the quantitative analysis of [Ca2+]i. © 2005 American Chemical Society.

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Citation Format(s)

Microfluidic selection and retention of a single cardiac myocyte, on-chip dye loading, cell contraction by chemical stimulation, and quantitative fluorescent analysis of intracellular calcium. / Li, Xiujun; Li, Paul C. H.

In: Analytical Chemistry, Vol. 77, No. 14, 15.07.2005, p. 4315-4322.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review