准确评估药物对活细胞 LDH 活性的影响：通过细胞核荧光染色和甲酰肼 LDH 活性双重测量活细胞数量。

IF 1.8 4区化学 Q3 CHEMISTRY, ANALYTICAL

Analytical Sciences Pub Date : 2024-07-20 DOI:10.1007/s44211-024-00631-6

Ryosuke Kaneko, Rui Hirakawa, Shoichi Hijii, Takeshi Mori, Yoshiki Katayama

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引用次数: 0

摘要

药物对乳酸脱氢酶（LDH）细胞内活性的影响是通过水溶性四氮唑（WST）来测定的。由于该测定通常是在死细胞存在的情况下进行的，因此无法评估活细胞的净活性。在此，我们报告了利用细胞核荧光染色对活细胞进行计数，从而测定活细胞细胞内 LDH 净活性的方法。通过使用深红色荧光染料，可在透明的 96 孔板上同时测量细胞核的荧光信号和 WST 的吸光度。我们发现，在有死细胞存在的情况下，传统的检测方法会高估药物对 LDH 活性的影响。

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Accurate evaluation of drug effect on the LDH activity of live cells: dual measurement of live cell number by fluorescent staining of nucleus and LDH activity by formazan

Effect of drugs on the intracellular activity of lactate dehydrogenase (LDH) has been measured by using water-soluble tetrazolium (WST). Because the assay is usually conducted in the presence of dead cells, net activity of live cells is not evaluated. Here, we reported the assay of the net intracellular LDH activity of live cells by counting the live cells using fluorescent staining of nucleus. By using a deep red fluorescent dye, dual measurements of fluorescence signal of nucleus and absorbance of WST could be conducted with transparent 96-well-plates. We found that conventional assay in the presence of dead cells overestimate the effect of drugs on the LDH activity.

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来源期刊

Analytical Sciences 化学-分析化学

CiteScore

2.90

自引率

18.80%

发文量

232

审稿时长

1 months

期刊介绍： Analytical Sciences is an international journal published monthly by The Japan Society for Analytical Chemistry. The journal publishes papers on all aspects of the theory and practice of analytical sciences, including fundamental and applied, inorganic and organic, wet chemical and instrumental methods. This publication is supported in part by the Grant-in-Aid for Publication of Scientific Research Result of the Japanese Ministry of Education, Culture, Sports, Science and Technology.