The invention discloses a nano particle for biological imaging and glioma targeting shown in a general formula I and a preparation method of the nano particle. The nano particle is formed by connecting tumor penetrating peptides and fluorescent carbon dots through maleimide-polyethylene glycol-amidogen succinimide succinic acid ester; and the value range of n is smaller than 85 and greater than 5 in the general formula I. The nano particle disclosed by the invention is targeted to brain glioma and then displays a good biological image; and applications in manners of intravenous injection, intramuscular injection and the like can be carried out in clinical. The general formula I is as shown in the description.