Scanless multitarget-matching multiphoton excitation fluorescence microscopy

Abstract

Using the combination of a re°ective blazed grating and a re°ective phase-only di®ractive spatial light modulator (SLM), scanless multitarget-matching multiphoton excitation°uorescence microscopy (SMTM-MPM) was achieved. The SLM shaped an incoming mode-locked, near-infrared Ti:sapphire laser beam into an excitation pattern with addressable shapes and sizes that matched the samples of interest in the¯eld of view. Temporal and spatial focusing were simultaneously realized by combining an objective lens and a blazed grating. The°uorescence signal from illuminated areas was recorded by a two-dimensional sCMOS camera. Compared with a conventional temporal focusing multiphoton microscope, our microscope achieved e®ective use of the laser power and decreased photodamage with higher axial resolution.

Aggregation-induced emission luminogen for in vivo three-photon fluorescence lifetime microscopic imaging

“…Thus, the images of these areas are lack of enough contrast. This could be improved by combining the 3PFM imaging with the°uorescence lifetime imaging microscopy (FLIM), which is suitable for noninvasive study of intracellular processes, 15,16 micro°uidic systems, 17 remote sensing, 18,19 lipid order problems in physical chemistry, 20 temperature sensing 21 and clinical medicine. 22 FLIM can provide a more sensitive and precise image based on the weak signals, compared with the tra-ditional°uorescence intensity imaging.…”