Quantitative imaging of mitochondrial redox conditions at the single-organelle level

Redox imaging

roGFP

Reactive oxygen species

Hypoxia

Aging

Mitochondria

2-photon microscopy

ΔΨm

mitochondrial membrane potential

ACSF

artificial cerebrospinal fluid

AMC

antimycin A

AR

aspect ratio

ATP

adenosine triphosphate

CI

capsula interna

cPu

caudoputamen

diamide

diazene dicarboxylic acid bis(N,N-dimethylamide)

DRP1

dynamin-related protein 1

DTT

1,4-dithio-DL-threitol

EUK-134

chloro[[2,2′-[1,2-ethanediylbis[(nitrilo-κN)methylidyne]] bis[6-methoxyphenolato-κO]]]-manganese

F740

fluorescence evoked by 740 nm excitation

F910

fluorescence evoked by 910 nm excitation

FCCP

carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone

FIS1

mitochondrial fission protein 1

FLIM

fluorescence lifetime imaging microscopy

GFP

green fluorescent protein

GSH / GSSG

reduced glutathione / oxidized glutathione disulfide

HIF-1α

hypoxia-inducible factor 1-alpha

H2O2

hydrogen peroxide

MFN1

mitofusin 1

MFN2

mitofusin 2

n

sample size

NA

numerical aperture

NADH

nicotinamide adenine dinucleotide

NADPH

nicotinamide adenine dinucleotide phosphate

NIR

near infrared

OPA1

OPA1 mitochondrial dynamin like GTPase

OxDroGFPm

relative degree of roGFPm oxidation

pd

postnatal day

PFA

paraformaldehyde

R

fluorescence ratio

Rox

ratio representing full roGFP oxidation

Rred

ratio representing full roGFP reduction

ROI

region of interest

roGFP

reduction/oxidation-sensitive green fluorescent protein

roGFPc

cytosolically expressed roGFP

roGFPm

mitochondria targeted roGFP

ROS

reactive oxygen species

tBHP

tert-butyl hydroperoxide

Thy1.2

thymocyte differentiation antigen 1

TPLSM

two photon laser scanning microscope