A microfabricated titration calorimeter having nW sensitivity is presented. The device is achieved by modifying a commercial, suspended-membrane, thin-film thermopile infrared sensor. Chemical reactions are studied by placing a 50.0 nL droplet of one reagent directly on the sensor and injecting nL droplets of a second reagent through a micropipette by means of a pressure-driven droplet injector with 1% reliability in volume delivery. External thermal noise is miminized by a two-layer thermal shielding system. Evaporation is prevented by positioning the micropipette through a tiny hole in a cover glass, sealed by a drop of oil. The device is calibrated using two acid-base reactions: H 2 SO 4 + HEPES buffer, and NaOH + HCl. The measured power sensitivity is 2.90(4) V/W, giving a detection limit of 22 nW. The 1/e time constant for a single injection is 1.1 s. The day-to-day power sensitivity is reproducible to ∼2%. A computational model of the sensor reproduces the power sensitivity within 10% and the time constant within 20%. For a 50 nL sample and 0.8 to 1.5 nL titrant injection volumes, the heat uncertainty of 44 nJ corresponds to a 3σ detection limit of 132 nJ, or the binding energy associated with 2.9 pM of IgG-protein A complex.Microcalorimetry is widely used to measure enthalpy changes in chemical reactions, biochemical processes, and phase transitions, with typical detection limits of μW or μJ. [1][2][3][4][5][6][7] In the last decade, micromachining techniques 8;9 have been used to produce calorimetric devices with greatly reduced sample volumes and detection capabilities in the nW range. [10][11][12][13] There is a growing realization that these advances could enlarge the role of calorimetry in drug discovery, 14 optimization of yeast metabolism 15
Experimental Section
Implementation of the CalorimeterThe S25 silicon-based thermopile sensor (Dexter Research Center, Inc., Dexter, MI) is shown in Figure 1. It has a twenty-junction thermopile with a Seebeck coefficient of 24 μV/K per junction. The thermopile is assembled under a thin (∼1.5 μm) membrane of SiO 2 /Si 3 N 4 . The metal tracks are directly below the membrane, and the sensing junctions are clustered at the center. Surrounding the free-standing membrane (the square shown in Figure 1D) is the aluminum heat sink that is attached to the porcelain body of the S25. The 0.5 mm well is formed by the membrane/heat sink at the bottom and the porcelain body of the sensor surrounding it. This well is modified to serve as our reaction chamber.The voltage signal generated by the S25 sensor is detected using an amplifier with a gain of 10 5 , then recorded by a PCI-6024E data acquisition board (National Instruments) in a desktop computer. A LabVIEW interface is used to control the experiment and display the readings.
Electrical Noise CharacterizationThe S25 has an intrinsic noise voltage of 19.4 nV/√Hz. We use a low-noise instrumentation amplifier and chopper-stabilized operational amplifier to minimize additional contributions to the intrinsic Johnso...