We have prepared novel poly(D,L-lactide-co-glycolide) (PLGA) lipid nanoparticles that covalently conjugate folic acid (FA) as well as indocyanine green (ICG) and also encapsulate resveratrol (RSV) (FA-RIPNPs), which are developed for simultaneous anticancer drug targeted delivery and fluorescence imaging. The FA-RIPNPs, with an average particle size of (92.8 ± 2.1) nm, were prepared by a facile self-assembly as well as nanoprecipitation method and showed excellent stability and biocompatibility characteristics. The FA-RIPNPs exhibited a RSV encapsulation efficiency of approximately (65.6 ± 4.7)% and a maximum release ratio of (78.2 ± 4.1)% at pH 5.0 and 37 ºC. Confocal fluorescence images showed that FA-RIPNPs may facilitate a high cellular uptake via FA receptor-mediated endocytosis... (RME). Furthermore, FA-RIPNPs (containing 50 μg/mL RSV) induced a (81.4 ± 2.1)% U87 cell inhibition rate via apoptosis, a value that proved to be higher than what has been shown for free RSV ((53.1 ± 1.1)%, equivalent RSV concentration). With a formulated polyethylene glycol (PEG) shell around the PLGA core, FA-RIPNPs prolonged blood circulation of free RSV as well as ICG, approximately increasing 6.96- and 39.4- fold (t1/2), respectively. As a NIR probe, in vivo fluorescence images indicated a highly efficient accumulation of FA-RIPNPs in tumorous tissue that proved to be approximately 2.8- and 12.6- fold higher than RIPNPs and free ICG, respectively. Intravenous injection of FA-RIPNPs into U87 tumor-bearing mice demonstrated the best tumor inhibition effect at all tested drugs, including free RSV and RIPNPs with no relapse, high biocompatibility and no significant systemic in vivo toxicity over the course of the treatment (one month). The results obtained demonstrate the versatility of the nanoparticles, featuring stable fluorescence and tumor targeting characteristics with promising future applications in cancer therapy.