13 March 2014 Non-contact measurement technique for enzymatic reaction of glucokinase
Author Affiliations +
Abstract
A non-contact infrared imaging-based measurement technique is applied to quantify the enzymatic reaction of glucokinase. The method is implemented by a long-wave (8-12 [μm]) infrared microbolometer imaging array and a germanium-based infrared optical vision system adjusted to the size of a small biological sample. The enzymatic reaction is carried out by the glucokinase enzyme, which is representative of the internal dynamics of the cell. Such reactions produce a spontaneous exothermal release of energy detected by the infrared imaging system as a non-contact measurement technique. It is shown by stoichiometry computations and infrared thermal resolution metrics that the infrared imaging system can detect the energy release at the [mK] range. This allows to quantify the spontaneity of the enzymatic reaction in a three dimensional (surface and time) single and noncontact real- time measurement. The camera is characterized for disclosing its sensibility, and the fixed pattern noise is compensated by a two point calibration method. On the other hand, the glucokinase enzyme is isolated from Pyrococcus furiosus. Therefore, the experiment is carried out by manual injection with graduated micropipettes using 40 [μl] of glucokinase at the surface of the substrate contained in an eppendorf tube. For recording, the infrared camera is adjusted in-focus at 25.4 [mm] from the superficial level of the substrate. The obtained values of energy release are 139 ± 22 [mK] at room temperature and 274 ± 22 [mK] for a bath temperature of 334 [K].
© (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Juan P. Staforelli, Juan P. Staforelli, María J. Gallardo, María J. Gallardo, Pablo Meza, Pablo Meza, Sergio Torres, Sergio Torres, Héctor Mella, Héctor Mella, Claudio Reyes, Claudio Reyes, "Non-contact measurement technique for enzymatic reaction of glucokinase", Proc. SPIE 8953, Optical Methods in Developmental Biology II, 89530L (13 March 2014); doi: 10.1117/12.2037394; https://doi.org/10.1117/12.2037394
PROCEEDINGS
9 PAGES


SHARE
Back to Top