13 March 2014 Non-contact measurement technique for enzymatic reaction of glucokinase
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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].
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Juan P. Staforelli, María J. Gallardo, Pablo Meza, Sergio Torres, Héctor Mella, 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
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