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

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

doi:10.1007/s44211-024-00631-6

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

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

Research output: Contribution to journal › Comment/debate › peer-review

Abstract

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. Graphical Abstract: (Figure presented.)

Original language	English
Pages (from-to)	2075-2080
Number of pages	6
Journal	analytical sciences
Volume	40
Issue number	11
DOIs	https://doi.org/10.1007/s44211-024-00631-6
Publication status	Published - Nov 2024

All Science Journal Classification (ASJC) codes

Analytical Chemistry

Access to Document

10.1007/s44211-024-00631-6

Cite this

@article{794bc836d21842c3abd93ac601724b39,

title = "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",

abstract = "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. Graphical Abstract: (Figure presented.)",

keywords = "Fluorometric and colorimetric dual-measurement, Lactate dehydrogenase, Live cell count, Tetrazolium",

author = "Ryosuke Kaneko and Rui Hirakawa and Shoichi Hijii and Takeshi Mori and Yoshiki Katayama",

note = "Publisher Copyright: {\textcopyright} The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry 2024.",

year = "2024",

month = nov,

doi = "10.1007/s44211-024-00631-6",

language = "English",

volume = "40",

pages = "2075--2080",

journal = "analytical sciences",

issn = "0910-6340",

publisher = "Springer International Publishing AG",

number = "11",

}

TY - JOUR

T1 - Accurate evaluation of drug effect on the LDH activity of live cells

T2 - dual measurement of live cell number by fluorescent staining of nucleus and LDH activity by formazan

AU - Kaneko, Ryosuke

AU - Hirakawa, Rui

AU - Hijii, Shoichi

AU - Mori, Takeshi

AU - Katayama, Yoshiki

N1 - Publisher Copyright: © The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry 2024.

PY - 2024/11

Y1 - 2024/11

N2 - 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. Graphical Abstract: (Figure presented.)

AB - 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. Graphical Abstract: (Figure presented.)

KW - Fluorometric and colorimetric dual-measurement

KW - Lactate dehydrogenase

KW - Live cell count

KW - Tetrazolium

UR - http://www.scopus.com/inward/record.url?scp=85199082436&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85199082436&partnerID=8YFLogxK

U2 - 10.1007/s44211-024-00631-6

DO - 10.1007/s44211-024-00631-6

M3 - Comment/debate

AN - SCOPUS:85199082436

SN - 0910-6340

VL - 40

SP - 2075

EP - 2080

JO - analytical sciences

JF - analytical sciences

IS - 11

ER -

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

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this