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首页> 外文期刊>Journal of bacteriology >Proton pumping and the internal pH of yeast cells, measured with pyranine introduced by electroporation.
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Proton pumping and the internal pH of yeast cells, measured with pyranine introduced by electroporation.

机译：质子泵入和酵母细胞的内部pH，通过电穿孔引入的吡喃测定。

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The internal pH of yeast cells was determined by measuring the fluorescence changes of pyranine (8-hydroxy-1,3,6-pyrene-trisulfonic acid), which was introduced into the cells by electroporation. This may be a suitable procedure for the following reasons. (i) Only minor changes in the physiological status of the cells seemed to be produced. (ii) The dye did not seem to leak at a significant rate from the cells. (iii) Different incubation conditions produced large fluorescence changes in the dye, which in general agree with present knowledge of the proton movements of the yeast cell under different conditions. (iv) Pyranine introduced by electroporation seemed to be located in the cytoplasm and to avoid the vacuole, and therefore it probably measured actual cytoplasmic pH. (v) Correction factors to obtain a more precise estimation of the internal pH are not difficult to apply, and the procedure may be useful for other yeasts and microorganisms, as well as for the introduction of other substances into cells. Values for the cytoplasmic pHs of yeast cells that were higher than those reported previously were obtained, probably because this fluorescent indicator did not seem to penetrate into the cell vacuole.
机译：通过测量通过电穿孔引入细胞中的吡喃（8-羟基-1,3,6-py-三磺酸）的荧光变化来确定酵母细胞的内部pH。由于以下原因，这可能是合适的过程。 （i）似乎只产生了细胞生理状态的微小变化。 （ii）染料似乎没有从细胞大量泄漏。 （iii）不同的孵育条件在染料中产生了很大的荧光变化，这通常与目前对酵母细胞在不同条件下质子运动的认识一致。 （iv）通过电穿孔引入的吡喃似乎位于细胞质中并避免液泡，因此它可能测量了实际的细胞质pH。 （v）不难应用校正因子以获得内部pH的更精确估计值，该程序可能对其他酵母和微生物以及向细胞中引入其他物质有用。获得的酵母细胞胞质pH值高于先前报道的值，可能是因为该荧光指示剂似乎未渗透到细胞液泡中。

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