How is cellular ATP measured?
Extracellular ATP is usually measured by adding luciferase and luciferin to cell cultures or to the cell supernatants. This is a sensitive and selective in vitro assay, but it is highly impractical for in vivo or real-time measurements.
Can you see ATP under a microscope?
Several methods to image ATP have been developed over the past many decades (Figure 2) (Table 1). Instead, methods to visualize ATP primarily rely on optical microscopy paired with molecular probes, which provide signal readouts that are specific for ATP.
What does ATP meter measure?
ATP bioluminescence meters, which measure the concentration of ATP as relative light units (RTU) in organic material and living cells [16], are widely used in food and beverage industries because of their ease of use and fast turn-around of results.
How does an ATP assay work?
Bioluminescent ATP assays take advantage of the firefly luciferase enzymatic reaction, which uses ATP from viable cells to generate photons of light. Viable cells are lysed to release the ATP for detection, and reagents containing firefly luciferase enzyme and substrate are added to catalyze a two-step reaction.
How is ATP measured in a living cell?
Method. ATP is a molecule found in and around living cells, and as such it gives a direct measure of biological concentration and health. ATP is quantified by measuring the light produced through its reaction with the naturally occurring firefly enzyme luciferase using a luminometer. The amount of light produced is directly proportional…
How are luciferase probes used to measure ATP in living cells?
Here, we describe optimized protocols for monitoring ATP concentrations in the cytosol, mitochondrial matrix and pericellular space in living cells via an overall procedure that requires an average of 3 d. In addition, we present a detailed protocol for the in vivo detection of extracellular ATP in mice using luciferase-transfected reporter cells.
Where is the ATP found in a water sample?
Within a water sample containing microorganisms, there are two types of ATP: Intracellular ATP – ATP contained within living biological cells. Extracellular ATP – ATP located outside of biological cells that has been released from dead or stressed organisms.
How are ATP levels measured in rat pancreas?
Ab83355 was used to determin ATP levels in rat pancreas islets as an ischemic marker to predict transplantation outcomes. We extracted ATP from fresh pancreas that have undergo different time of cold ischemia : 0, 2, 4, 6, 8 and 10h and in situ.