In this study, we reported on the synthesis and biological evaluation of radiolabeled fluorescent dye conjugated bovine serum albumin nanoparticles within the size range 190–210nm. The bovine serum albumin nanoparticles (BSANPs) were prepared using a desolvation method, and chemical cross-linking was performed using gluteraldehyde. Furthermore, pheophorbide-a (PH-A) was loaded on the BSANPs. The results obtained from dynamic light scattering and electron microscopy have proved that nanoparticles are highly monodisperse and near-spherical shaped. The photo-physical properties of the PH-A-BSANPs were obtained using the spectrophotometric techniques. According to the results, PH-A and BSANPs show high non-covalent interaction. PH-A loaded nanoparticles were labeled with ^99mTc and the radio-labeling efficiency was determined as 90±1.2%. Biodistribution studies of ^99mTc labeled PH-A-BSANPs and PH-A were carried out using female Albino Wistar rats, and ^99mTc-PH-A-BSANPs showed a significantly higher uptake in the breast and uterus than ^99mTc-PH-A. Cell culture study was carried out in MCF-7 cell line (human breast adenocarcinoma cell line). According to the cell culture studies, ^99mTc-PH-A-BSANPs showed a higher uptake than ^99mTc-PH-A. Moreover, PH-A-BSANPs demonstrated good photo-physical properties and BSANPs increased the uptake of PH-A on to the MCF-7 cell line. These results confirm that ^99mTc labeled PH-A-BSANPs could be utilized for radioimaging.