A graft copolymer composed of poly(ε-caprolactone) as a hydrophobic chain and poly(N-(2-hydroxypropyl)methacrylamide) as a hydrophilic segment with an internal reference probe for oxygen sensing was prepared. Highly efficient platinum(II)meso-tetra(pentafluorophenyl)porphine (PtTFPP) was used as a typical oxygen probe. Although PtTFPP was highly efficient, it is hydrophobic, limiting its direct application for oxygen sensing in biological conditions. By using the amphiphilic graft copolymer, PTTFPP was successfully incorporated into the micelles formed by this polymer to not only enable the application of PtTFPP for biosensing in medium but also retain its high efficiency. The quantum efficiency of PtTFPP in the micelle solution under nitrogen can reach as high as 0.20. The PtTFPP-containing micelles were tested for high throughput cell respiration and bacteria detection. The influence of an antibiotic, antimycin as a representative example, on cell respiration was also studied. For achieving high accurate sensing, an internal built-in reference probe with aggregation induced emission properties were also polymerized in the graft copolymer. This study demonstrated that the graft polymer approach is a good way to enable highly efficient and hydrophobic oxygen probes for biosensing.