Optical ATP biosensor for extracellular ATP measurement

• Published in: Biosensors & bioelectronics 2013-05, Vol.43, p.355-361
• Main Authors: Wang, C., Huang, C.-Y.C., Lin, W-C
• Format: Article
• Language: English
• Subjects: adenosine triphosphate; Adenosine Triphosphate - analysis; Adenosine Triphosphate - chemistry; ATP; Biological and medical sciences; Biosensing Techniques - instrumentation; Biosensor; Biosensors; Biotechnology; coatings; Compensation methods; Equipment Design; Equipment Failure Analysis; Extracellular Fluid - chemistry; Fiber Optic Technology - instrumentation; Fundamental and applied biological sciences. Psychology; glycerol kinase; intervertebral disks; Methods. Procedures. Technologies; Optical; Optical Devices; oxidation; oxygen; Oxygen - analysis; Oxygen - chemistry; receptors; Refractometry - instrumentation; ruthenium; Ruthenium complex; swine; Various methods and equipments
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2012.12.027

Extracellular Adenosine-5′-triphosphate (ATP) is an important multi-functional molecule which can mediate numerous physiological activities by activating purinergic P2 receptors. The objective of this study was to develop a novel optical ATP sensor for in-situ extracellular ATP measurement in biological tissues. The optical ATP sensor was made by applying two layers of sol–gel coating to the end of an optical fiber probe end. The first layer contained ruthenium complex for sensing changes in oxygen concentration which resulted from oxidation of ATP by glycerol kinase and glycerol 3-phosphate oxidase entrapped in the second layer. It was demonstrated that the optical ATP sensor was capable of detecting ATP concentration at a broad range of 10−3mM to 1.5mM.A compensation method was established to enable the optical sensor to determine ATP concentration at different oxygen levels. This study also demonstrated the capability of ATP sensor to measure extracellular ATP content in biological tissues (i.e., porcine intervertebral disc). In addition, it was shown that the optical ATP sensor was not affected by pH and derivatives of extracellular ATP. Therefore, the newly developed optical ATP sensor is a good option for in-situ extracellular ATP measurement. ► A novel ATP biosensor developed based on optical oxygen sensing technique and enzymatic ATP breakdown. ► The new biosensor can detect the ATP concentrations ranged from 10−3mM to 1.5mM. ► A compensation method for different oxygen levels was proposed and tested. ► An in situ measurement of extracellular ATP level in intervertebral disc was presented.