Optical nanoprobes for biosensing applications have gained widespread attention for recognition of a variety of biorelevant ions or molecules present in human body. In the present chapter, the nanoprobes are classified into four categories: zero, one, two, and three-dimensional nanomaterials and herein the nanomaterials include carbon quantum dots (CQDs), graphene quantum dots (GQDs), semiconductor inorganic quantum dots (IQDs), polymer dots (PDs), gold nanoparticles (AuNPs), silver nanoparticles (AgNPs), upconversion nanoparticles (UCNPs), magnetic nanoparticles (MNPs) for 0-D, carbon nanotubes (CNTs), gold nanorods (AuNRs), silicon nanowires (SiNWs), nanofibers and nanoribbons for 1-D, MnO 2 nanosheets, 2-D MOF nanosheets, graphitic carbon nitride, graphene oxide (GO), reduced graphene oxide (rGO) for 2-D and hybrid nanoflowers (HNFs) and 3-D MOFs for 3-D nanomaterials. The chapter briefly covers diverse nanomaterials and nanocomposites and their applications in chromo-fluorgenic biosensing applications. Furthermore, the applications of the nanoprobes for targeted analytes recognition from complicated biological samples as well as in intracellular imaging have also been discussed. Finally, the loopholes of the present research and future directions of the research are also enlightened to keep an ample footstep in the domain of medical diagnosis.