Bioluminescent calcium fluorapatite nanocrystals (TCaFAp:Eu) with controlled morphology were fabricated through an easy one-step method and found to have excellent water dispersibility and biocompatibility. Further modification of conjugation with both optical imaging and cell targeting capabilities provided useful nanoprobes for highly sensitive biorecognition applications, which was realized by grafting folic acid onto the surface of TCaFAp:Eu. The uptake of designed TCaFAp:Eu-FA by HepG2 cancer cells was verified by confocal laser scanning microscopy after 30min, whereas the uptake of TCaFAp:Eu-FA by MCF-7 cells did not work, indicating that the prepared nanoprobe effectively targeted the imaging of cancer cells with over-expression of folate receptor.