A sensitive and reliable bimodal electrochemiluminescent (ECL) system based on CdTe quantum dots (QDs) and luminol as double luminophores is constructed. CdTe QDs tagged with the aptamer (CdTe-Apt 2) of cancer cells are used as the detection signals, while luminol molecules are used as internal standards. The electrodeposited polyaniline-based conducting polymer hydrogel (CPH) on the electrode surfaces improves the biocompatibility and conductivity of the sensing interfaces effectively. Furthermore, electron transfer is probably much easier when luminol and coreactant potassium persulfate (K₂S₂O₈) are immobilized in the CPH in comparison to that in solution. Cancer cells are captured to the electrode surface by another aptamer linked to the Au nanoparticles immobilized in the CPH through Au–S bonds. In the developed bimodal ECL system, an internal standard method is used to quantify cancer cells by comparing the differences in sensitivity of the double-peak ECL signals with that of target analytes. The internal standard method of ECL strategy can provide very accurate detection results in a complex environment because interferences in the system can be eliminated through the self-calibration of two emission spectra. A linear relation is found on the basis of a plot of the ΔECL_CdTe/ΔECL_luminol against the concentration of cancer cells within 100–6500 cells mL^–1 under optimized conditions. The developed ratiometric ECL cytosensor with internal standard can significantly improve the accuracy and reliability of cell assays in complex biological media, demonstrating promising applications in healthcare monitoring and clinical diagnostics.