Evaluation of the cell health status is critical for drug screening and cell physiological activity investigations. The existing cell health assessment methods are solely devoted to the study of cell vitality or viability, leading to an incomplete evaluation. Herein, we report a convenient and robust method for the joint assessment of cell viability and vitality based on electric cell-substrate impedance sensing (ECIS) supplied with an environmental temperature control. The static value of electric cell-substrate impedance reflects the survival rate of cells, while the temperature tolerance of cells demonstrates the cell vitality. It was found that the cell vitality evaluated by the temperature tolerance of cells was independent of the initial cell numbers, rendering the proposed method easy to utilize in various applications. We compared the temperature tolerance ECIS method with the traditional trypan blue staining method, the methyl thiazolyl tetrazolium assay, and the direct impedance sensing method for joint evaluation of cell viability and vitality in drug screening. The temperature tolerance ECIS method showed comparable results but with a simpler protocol, faster results, and less dependence on the sample conditions. By providing both information on cell viability and cell vitality, the proposed temperature tolerance ECIS method would pave the way in building a simple and robust sensing system for cell health evaluation.