As a new kind of advanced nonlinear imaging approach, two-photon fluorescence microscopy technology is wildly used in the field of live cell and tissue imaging, especially focusing on long-term dynamic three-dimensional cell imaging. This paper firstly presents the principle and characteristic of two-photon fluorescence microscopy. Then, the paper focuses on the three key aspects of the viability of the specimen, sensitivity of detection, as well as the speed of acquisition. In the end, the future prospect of development and application of two-phonon imaging technology are predicted.
Diagnostic Imaging ; Microscopy, Fluorescence, Multiphoton ; methods