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首页> 外文期刊>Journal of Microbiological Methods >Assessment of FUN-1 vital dye staining: Yeast with a block in the vacuolar sorting pathway have impaired ability to form CIVS when stained with FUN-1 fluorescent dye
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Assessment of FUN-1 vital dye staining: Yeast with a block in the vacuolar sorting pathway have impaired ability to form CIVS when stained with FUN-1 fluorescent dye

机译：FUN-1活性染料染色的评估：在液泡分选途径中受阻的酵母在用FUN-1荧光染料染色时形成CIVS的能力受损

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摘要

FUN-1 [2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methyli d ene)-1-phenylquinolinium iodide] is a fluorescent dye used in studies of yeast and other fungi to monitor cell viability in the research lab and to assay for active fungal infection in the clinical setting. When the plasma membrane is intact, fungal cells internalize FUN-1 and the dye is seen as diffuse green cytosolic fluorescence. FUN-1 is then transported to the vacuole in metabolically active wild type cells and subsequently is compacted into fluorescent red cylindrical intravacuolar structures (CIVS) by an unknown transport pathway. This dye is used to determine yeast viability, as only live cells form CIVS. However, in live Saccharomyces cerevisiae with impaired protein sorting to the yeast vacuole, we report decreased to no CIVS formation, depending on the cellular location of the block in the sorting pathway. Cells with a block in vesicle-mediated transport from the Golgi to prevacuolar compartment (PVC) or with a block in recycling from the PVC to the Golgi demonstrate a substantial impairment in CIVS formation. Instead, the FUN-1 dye is seen either in small punctate structures under fluorescence or as diffuse red cytosol under white light. Thus, researchers using FUN-1 should be cognizant of the limitations of this procedure in determining cell viability as there are viable yeast mutants with impaired CIVS formation.
机译：FUN-1 [2-氯-4-（2,3-二氢-3-甲基-（苯并-1,3-噻唑-2-基）-甲基碘）-1-苯基喹啉碘化物]是一种荧光染料在酵母和其他真菌的研究中，负责监测研究实验室中的细胞活力，并在临床环境中检测活性真菌感染。当质膜完好无损时，真菌细胞将FUN-1内在化，并且该染料被视为弥漫性绿色胞质荧光。然后将FUN-1转运至具有代谢活性的野生型细胞中的液泡中，然后通过未知的转运途径将其压实为荧光红色圆柱状真空内结构（CIVS）。该染料用于确定酵母的活力，因为只有活细胞才能形成CIVS。然而，在活酵母中，对酵母液泡的蛋白质分选受损，我们报道其下降至没有CIVS形成，这取决于分选途径中该嵌段的细胞位置。从高尔基体到高泡前腔室（PVC）的囊泡介导运输受阻的细胞，或从PVC到高尔基体的再循环中受阻的细胞均表现出CIVS形成的实质性损害。取而代之的是，FUN-1染料要么在荧光的小点状结构中看到，要么在白光下看到的是弥漫性红色胞浆。因此，使用FUN-1的研究人员应该认识到此程序在确定细胞活力方面的局限性，因为存在存活的酵母突变体，其CIVS形成受损。

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