Non-contact Measurement Technique for Enzymatic Reaction of Glucokinase

Staforelli, JP; Gallardo, MJ; Meza, P; Torres, S.; Mella, H; Reyes, C.

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 [mu m]) 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 [mu] 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].

Más información

Título según WOS: Non-contact Measurement Technique for Enzymatic Reaction of Glucokinase
Título según SCOPUS: Non-contact measurement technique for enzymatic reaction of glucokinase
Título de la Revista: COMPUTATIONAL OPTICS 2024
Volumen: 8953
Editorial: SPIE-INT SOC OPTICAL ENGINEERING
Fecha de publicación: 2014
Idioma: English
DOI:

10.1117/12.2037394

Notas: ISI, SCOPUS