Extrinsic Fabry-Perot interferometry for noncontact temperature control of nanoliter-volume enzymatic reactions in...

Optical fiber extrinsic Fabry-Perot interferometry (EFPI) was investigated as a noncontact temperature sensor and utilized for regulating the temperature of small-volume solutions in microchips. Interference pattern analysis determined the optical path lengths (OPL) associated with reflections from various surfaces on or in the microchip, in particular, from gold sputtered on the bottom of a microchannel. Since OPL is directly proportional to refractive index, which is dependent on solution temperature, the EFPI sensor was capable of noncontact monitoring of solution temperature simply from alterations in the measured path length. Calibration of the sensor against a thermocouple was performed while heating the microchip in a noncontact manner with an IR lamp. The combination of EFPI temperature sensor, IR-mediated heating, and air cooling allowed a fully noncontact system for small-volume temperature control in microchip structures, and its utility was illustrated by optimal digestion of DNA by a temperature-dependent restriction endonuclease in 320 nL. The functionality and simplicity of the microchip EFPI temperature sensor was enhanced by replacing the prebonding sputtered gold with a tunable, chemically plated semireflective silver coating created in situ after chip fabrication. This provided an 8-fold improvement in the lowest detectable temperature change (deltaT = 0.1 degrees C), facilitated primarily by enhanced reflection from both the bottom and top surfaces of the microchannel. This approach for controlling micro- and nanoscale reactions--with heating, cooling, and temperature control being carried out in a completely noncontact fashion--provides an accurate and sensitive method for executing chemical and biochemical reactions in microchips.