In this study, a microfluidic-based physiometer capable of measuring whole blood viscosity, hematocrit, and red blood cell (RBC) deformability on a chip is introduced. The physiometer consists of two major parts: a hydrodynamic component for whole blood viscosity measurement and an electronic component for hematocrit and RBC deformability measurement. In the hydrodynamic component, the whole blood is infused with phosphate buffered saline as a reference fluid for estimation of the whole blood viscosity. At a given flow rate, ten sets of whole blood viscosity readings are successfully obtained over a wide range of shear rates; this is achieved via a series of geometrically optimized microchannel arrays. In the electronic component, analysis of the whole blood impedance spectrum under flowing conditions reveals the electrical characteristics of the blood: the cytoplasm resistance (Rcytoplsm), plasma resistance (Rplasma), and RBC membrane capacitance (constant phase element). The hematocrit is estimated from Rcytoplsm and Rplasma, while the RBC deformation index is determined from the membrane capacitance change of the RBC. Each unique function is experimentally demonstrated and compared to the corresponding gold standard method. The whole blood viscosity measured using the physiometer is 0.8 ± 1.4% in normalized difference compared to that using a rotational cone-and-plate viscometer. For the hematocrit measurement, the coefficient of variation for the physiometer ranges from 0.3 to 1.2% which is lower than the one obtained from centrifugation. In the deformability measurement, there is a strong linear correlation (R2 = 0.97) between the deformation index acquired by image processing and the change in the membrane capacitance acquired by using the physiometer. The effects of the hematocrit and RBC deformability on the whole blood viscosity are also demonstrated. For simultaneous and reliable measurement on a chip, a physiometer equipped with a temperature-control system is prepared. Lab-made software enables the measurement of the three target indices and the temperature control in an automated manner. By using this system, the temperature is controlled to 36.9 ± 0.2 °C which greatly matches with the target temperature (37.0 °C) and it is varied from 25 °C to 43 °C. The developed physiometer is potentially applicable for a comprehensive analysis of biophysical indices in whole blood.