A high precision Levenberg-Marquardt (LM) iterative algorithm for the analysis of fluorescence lifetime imaging microscopy (FLIM) data is reported. The performance of this method was tested on the time-resolved fluorescence intensity images of fluorescence lifetime standard dyes and real biological images. The method is applicable for various fluorescence decay functions and has better estimation precision based on iterative nonlinear least squares minimization algorithm. It indicates that the LM iterative technique for FLIM analysis represents a precise and versatile method that enables practical applications of FLIM in biology, biochemistry, biophysics, and medical diagnosis.