An adaptive TFI reconstructing method includes converting a diffusion equation into a linear equation through a finite element method; constructing a linear relation between unknown fluorescent light source distribution and a surface fluorescence measurement value; calculating a current regularization parameter and selecting an element with the largest absolute value in a difference correlation coefficient into a support set I; extracting all the current elements in the updated support set from corresponding lines in a matrix A, constituting a matrix, and obtaining a next-step searching direction; calculating the length of the next step and updating the support set; obtaining a next-step result according to the solved searching direction and step length iteration, and updating the regularization parameter; and determining whether stopping requirements are satisfied, if yes, finishing the reconstructing process, or otherwise, turning to the fourth step. According to the method, the regularization parameter is not needed to predict but determined adaptively during the reconstructing method, the reconstructing robustness is improved through adaptive regularization parameter selection, and the reconstructing efficiency is improved greatly.