Minimizing ATP depletion by oxygen scavengers for single-molecule fluorescence imaging in live cells.

ABSTRACT: The stability of organic dyes against photobleaching is critical in single-molecule tracking and localization microscopy. Since oxygen accelerates photobleaching of most organic dyes, glucose oxidase is commonly used to slow dye photobleaching by depleting oxygen. As demonstrated here, pyranose-2-oxidase slows bleaching of Alexa647 dye by ∼20-fold. However, oxygen deprivation may pose severe problems for live cells by reducing mitochondrial oxidative phosphorylation and ATP production. We formulate a method to sustain intracellular ATP levels in the presence of oxygen scavengers. Supplementation with metabolic intermediates including glyceraldehyde, glutamine, and α-ketoisocaproate maintained the intracellular ATP level for at least 10 min by balancing between FADH₂ and NADH des

SUBMITTER: Jung SR 

PROVIDER: S-EPMC6016803 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature