Simple and disposable monitoring of blood is usually the best solution for early clinical diagnosis and home self-inspection of the chronic patients. Herein, a simple point-of-care (POC) device which called molecularly imprinted membrane modified gel colorimetric device (MIMGCD) was prepared for whole blood cholesterol colorimetric detection at the first time. The convenient detection principle of this device rely on molecularly imprinted membranes for specifically separating cholesterol from whole blood firstly, following the gold bipyramids (GBPs) agarose gel system is react with the cholesterol oxidation to product hydrogen peroxide (H2O2), and the cholesterol will be quantified based on the color change. Under optimal conditions, the analytical performance of the proposed device yielded a linear range of 315.8–6000.0 μM and detection limit of 94.7 μM with 6.89% RSD for cholesterol, which can meet the needs of the detection of normal cholesterol content in the human body. Compared with the traditional whole blood detection methods, no complex sample preparation steps or precision instruments are required, endowing MIMGCD with the merits of easy to operate and low-cost. In addition, the multicolor variation of GBPs in the device allow a colorimetric card-like detection mechanism, which can be used for home self-inspection. This device has the potential to be utilized in clinical and home POC testing application for whole blood biomolecule analysis, thereby facilitating the whole blood screening and long-term monitoring in non-specialized laboratory infrastructure.