Cell imaging and bioactive molecules detection in living cells with long-lived lanthanide-based probes based on time-resolved fluorescence microscope, recently, have emerged as promising tools for the diagnosis of human diseases. In this research, a series of amphiphilic lanthanide (Ln = YbIII, ErIII) porphyrinate with high NIR emission quantum yield and specific cell location were constructed. The ytterbium complex (Yb-Rh) was found not only impressive NIR emission with linear and nonlinear excitations in water but also mitochondria specific in HeLa cells. Moreover, an ytterbium complex (Yb-N) was found that it can recognize in-vitro anionic phospholipids confirmed with the investigation of their performance in five cancer cell lines and three normal cell lines. The primary research goals are directed toward understanding basic photoproperties of the porphyrinate lanthanide complexes for the construction of long-lived NIR emissive probes to detect specific bioactive analytes in living cells.