FLIM

Super-resolution fluorescence microscopy by stepwise optical saturation

Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the super-resolution microscopy …

Description of deep saturated excitation multiphoton microscopy for super-resolution imaging

Here we recount the standard two-level model that describes saturated excitation (SAX) in multiphoton microscopy (MPM), a new technique for super-resolution fluorescence microscopy in scattering tissue, which requires no special chemistry and only …

Doubling the sensitivity of multiphoton frequency-domain fluorescence lifetime images

We demonstrate super-sensitive multiphoton frequency-domain fluorescence lifetime images using the recently developed DC&1ω method by showing a two-fold improvement in imaging sensitivity compared to the conventional 1ω phase fluorometry.

Silica-coated ruthenium-complex nanoprobes for two-photon oxygen microscopy in biological media

Multiphoton microscopy (MPM) allows for three-dimensional in vivo microscopy in scattering tissue with submicron resolution and high signal-to-noise ratio. MPM combined with fluorescence lifetime measurements further enables quantitative imaging of …

Super-sensitivity and super-resolution multiphoton fluorescence lifetime imaging microscopy

To fundamentally understand biological and medical phenomena such as aging and diseases, we need the information of the microenvironment surrounding living cells. We propose the multiphoton fluorescence lifetime imaging microscopy (MPM-FLIM) platform …

Saturation-compensated measurements for fluorescence lifetime imaging microscopy

Fluorophore saturation is the key factor limiting the speed and excitation range of fluorescence lifetime imaging microscopy (FLIM). For example, fluorophore saturation causes incorrect lifetime measurements when using conventional frequency-domain …

Super-sensitivity multiphoton frequency-domain fluorescence lifetime imaging microscopy

We present a series of experiments that demonstrate a super-sensitive chemical imaging technique based on multiphoton frequency-domain fluorescence lifetime imaging microscopy (MPM-FD-FLIM) that shows a 2× improvement in imaging speed compared to the …

Investigation of signal-to-noise ratio in frequency-domain multiphoton fluorescence lifetime imaging microscopy

Multiphoton microscopy (MPM) combined with fluorescence lifetime imaging microscopy (FLIM) has enabled three-dimensional quantitative molecular microscopy in vivo. The signal-to-noise ratio (SNR), and thus the imaging rate of MPM-FLIM, which is …

Highly stable two-photon oxygen imaging probe based on a ruthenium-complex encapsulated in a silica-coated nanomicelle

A nanomicelle-based ruthenium-complex oxygen imaging probe is coated with a silica shell. The biostability of the silica-coated probe is improved by a factor of 4, while the oxygen-sensitivity is reduced by a factor of 3.

Theoretical analysis of the signal-to-noise ratio of two-photon oxygen imaging probes

Signal-to-noise ratio performance of optical chemical sensors in the presence of phosphorescence saturation is described using Poisson statistics and Stern-Volmer kinetics. The described framework can be applied across a wide range of sensitivites …