Two-photon active fluorescent indicator for detecting NADH dynamics in live cells and tumor tissue

Arup Podder; Natesan Thirumalaivasan; Yu Kai Chao; Prasanna Kukutla; Shu Pao Wu; Sankarprasad Bhuniya

doi:10.1016/j.snb.2020.128637

Two-photon active fluorescent indicator for detecting NADH dynamics in live cells and tumor tissue

Arup Podder, Natesan Thirumalaivasan, Yu Kai Chao, Prasanna Kukutla, Shu Pao Wu^*, Sankarprasad Bhuniya

^*Corresponding author for this work

Department of Applied Chemistry

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

We have developed a two-photon fluorescent indicator (MC_2P) that participates in a redox reaction with NADH to provide a fluorescence signal at 460 nm. The quantum yield of the indicator MC_2P has increased about 26-fold in the presence of NADH. High two-photon efficiency (GM = 80.94) of the indicator MC_2P has considered it as a unique device for assessing NADH expression in live cells without interference from native fluorogenic entities. It rapidly internalized into live cells and showed a considerable fluorescence signal in cancer cells than the immune cells (RAW 264.7). The indicator MC_2P has provided information on the fluctuation of NADH levels depending on the variation of metabolites in the glycolysis pathway. Further, we noticed an increment of NADH level in mitochondrial respiratory cascade by inhibiting NADH-ubiquinone oxo reductase (complex I) using rotenone. On the contrary, uncoupler (CCCP) induced depletion of NADH. The dynamic expression of NAD(P)H has increased for counterbalancing mitochondrial redox potential during ROS (H₂O₂) suppression by GSH. Finally, in an vivo experiment, the indicator MC_2P has distinguished colon tumor (HT-29 cells) tissue from healthy tissue by providing two-photon fluorescence images.

Original language	English
Article number	128637
Journal	Sensors and Actuators, B: Chemical
Volume	324
DOIs	https://doi.org/10.1016/j.snb.2020.128637
State	Published - 1 Dec 2020

Keywords

Fluorescence
NADH
Redox potential
Tumor tissue
Two-photon

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10.1016/j.snb.2020.128637

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title = "Two-photon active fluorescent indicator for detecting NADH dynamics in live cells and tumor tissue",

abstract = "We have developed a two-photon fluorescent indicator (MC2P) that participates in a redox reaction with NADH to provide a fluorescence signal at 460 nm. The quantum yield of the indicator MC2P has increased about 26-fold in the presence of NADH. High two-photon efficiency (GM = 80.94) of the indicator MC2P has considered it as a unique device for assessing NADH expression in live cells without interference from native fluorogenic entities. It rapidly internalized into live cells and showed a considerable fluorescence signal in cancer cells than the immune cells (RAW 264.7). The indicator MC2P has provided information on the fluctuation of NADH levels depending on the variation of metabolites in the glycolysis pathway. Further, we noticed an increment of NADH level in mitochondrial respiratory cascade by inhibiting NADH-ubiquinone oxo reductase (complex I) using rotenone. On the contrary, uncoupler (CCCP) induced depletion of NADH. The dynamic expression of NAD(P)H has increased for counterbalancing mitochondrial redox potential during ROS (H2O2) suppression by GSH. Finally, in an vivo experiment, the indicator MC2P has distinguished colon tumor (HT-29 cells) tissue from healthy tissue by providing two-photon fluorescence images.",

keywords = "Fluorescence, NADH, Redox potential, Tumor tissue, Two-photon",

author = "Arup Podder and Natesan Thirumalaivasan and Chao, {Yu Kai} and Prasanna Kukutla and Wu, {Shu Pao} and Sankarprasad Bhuniya",

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doi = "10.1016/j.snb.2020.128637",

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AU - Podder, Arup

AU - Thirumalaivasan, Natesan

AU - Chao, Yu Kai

AU - Kukutla, Prasanna

AU - Wu, Shu Pao

AU - Bhuniya, Sankarprasad

PY - 2020/12/1

Y1 - 2020/12/1

N2 - We have developed a two-photon fluorescent indicator (MC2P) that participates in a redox reaction with NADH to provide a fluorescence signal at 460 nm. The quantum yield of the indicator MC2P has increased about 26-fold in the presence of NADH. High two-photon efficiency (GM = 80.94) of the indicator MC2P has considered it as a unique device for assessing NADH expression in live cells without interference from native fluorogenic entities. It rapidly internalized into live cells and showed a considerable fluorescence signal in cancer cells than the immune cells (RAW 264.7). The indicator MC2P has provided information on the fluctuation of NADH levels depending on the variation of metabolites in the glycolysis pathway. Further, we noticed an increment of NADH level in mitochondrial respiratory cascade by inhibiting NADH-ubiquinone oxo reductase (complex I) using rotenone. On the contrary, uncoupler (CCCP) induced depletion of NADH. The dynamic expression of NAD(P)H has increased for counterbalancing mitochondrial redox potential during ROS (H2O2) suppression by GSH. Finally, in an vivo experiment, the indicator MC2P has distinguished colon tumor (HT-29 cells) tissue from healthy tissue by providing two-photon fluorescence images.

AB - We have developed a two-photon fluorescent indicator (MC2P) that participates in a redox reaction with NADH to provide a fluorescence signal at 460 nm. The quantum yield of the indicator MC2P has increased about 26-fold in the presence of NADH. High two-photon efficiency (GM = 80.94) of the indicator MC2P has considered it as a unique device for assessing NADH expression in live cells without interference from native fluorogenic entities. It rapidly internalized into live cells and showed a considerable fluorescence signal in cancer cells than the immune cells (RAW 264.7). The indicator MC2P has provided information on the fluctuation of NADH levels depending on the variation of metabolites in the glycolysis pathway. Further, we noticed an increment of NADH level in mitochondrial respiratory cascade by inhibiting NADH-ubiquinone oxo reductase (complex I) using rotenone. On the contrary, uncoupler (CCCP) induced depletion of NADH. The dynamic expression of NAD(P)H has increased for counterbalancing mitochondrial redox potential during ROS (H2O2) suppression by GSH. Finally, in an vivo experiment, the indicator MC2P has distinguished colon tumor (HT-29 cells) tissue from healthy tissue by providing two-photon fluorescence images.

KW - Fluorescence

KW - NADH

KW - Redox potential

KW - Tumor tissue

KW - Two-photon

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Two-photon active fluorescent indicator for detecting NADH dynamics in live cells and tumor tissue

Abstract

Keywords

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