Journal ArticleDOI
Compact multiphoton/single photon laser scanning microscope for spectral imaging and fluorescence lifetime imaging
TLDR
To expand the features of the compact LSM, a time-correlated single photon counting unit as well as a Sagnac interferometer have been added to realize fluorescence lifetime imaging (FLIM) and spectral imaging.Abstract:
An inverted fluorescence microscope was upgraded into a compact three-dimensional laser scanning microscope (LSM) of 65 x 62 x 48 cm dimensions by means of a fast kHz galvoscanner unit, a piezodriven z-stage, and a picosecond (ps) 50 MHz laser diode at 405 nm. In addition, compact turn-key near infrared femtosecond lasers have been employed to perform multiphoton fluorescence and second harmonic generation (SHG) microscopy. To expand the features of the compact LSM, a time-correlated single photon counting unit as well as a Sagnac interferometer have been added to realize fluorescence lifetime imaging (FLIM) and spectral imaging. Using this unique five-dimensional microscope, TauMap, single-photon excited (SPE), and two-photon excited (TPE) cellular fluorescence as well as intratissue autofluorescence of water plant leaves have been investigated with submicron spatial resolution, < 270 ps temporal resolution, and 10 nm spectral resolution.read more
Citations
More filters
Journal ArticleDOI
Spectrally Resolved Multiphoton Imaging of In Vivo and Excised Mouse Skin Tissues
Jonathan Alambra Palero,Henriëtte S. de Bruijn,Angelique van der Ploeg-van den Heuvel,Henricus J. C. M. Sterenborg,Hans C. Gerritsen +4 more
TL;DR: Spectral imaging provided a wealth of information not easily obtainable with present conventional multiphoton imaging systems and suggested that the origin of the narrowband emission was collagen Raman peaks.
Journal Article
Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope
Martin Straub,Stefan W. Hell +1 more
TL;DR: The combination of pulsed-mode excitation multifocal multiphoton microscopy with a high-repetition, time-gated intensified CCD camera enables efficient three-dimensional (3D) fluorescence lifetime imaging as discussed by the authors.
Journal ArticleDOI
Multiphoton fluorescence lifetime imaging of 3D-stem cell spheroids during differentiation.
TL;DR: Time‐resolved multiphoton autofluorescence imaging microscopes may become a promising tool for marker‐free stem‐cell characterization and cell sorting.
Journal Article
Comparison of three-dimensional imaging properties between two-photon and single-photon fluorescence microscopy
Min Gu,Colin J. R. Sheppard +1 more
TL;DR: In this article, the imaging performance in single-photon (l-p) and two-p (2p) fluorescence microscopy is compared in terms of the three-dimensional (3D) point spread function and the 3-D optical transfer function.
Journal ArticleDOI
In vivo nonlinear spectral imaging in mouse skin
Jonathan Alambra Palero,H.S. de Bruijn,A. van der Ploeg van den Heuvel,Henricus J. C. M. Sterenborg,Hans C. Gerritsen +4 more
TL;DR: The results show that linking nonlinear deep-tissue imaging microscopy with autofluorescence spectroscopy has the potential to provide important information for the diagnosis of skin tissues.
References
More filters
Journal ArticleDOI
Two-Photon Laser Scanning Fluorescence Microscopy
TL;DR: The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
Journal ArticleDOI
Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm
Chris Xu,Watt W. Webb +1 more
TL;DR: In this paper, the two-photon fluorescence excitation (TPE) spectra were measured for 11 common molecular fluorophores in the excitation wavelength range 690 nm < λ < 1050 nm.
Journal ArticleDOI
Multicolor Spectral Karyotyping of Human Chromosomes
Evelin Schröck,S du Manoir,T Veldman,B Schoell,Johannes Wienberg,Malcolm A. Ferguson-Smith,Yi Ning,David H. Ledbetter,Irit Bar-Am,D. Soenksen,Yuval Garini,Thomas Ried +11 more
TL;DR: Whole-genome scanning by spectral karyotyping allowed instantaneous visualization of defined emission spectra for each human chromosome after fluorescence in situ hybridization, and all human chromosomes were simultaneously identified.
Journal ArticleDOI
Two-photon fluorescence excitation cross sections of biomolecular probes from 690 to 960 nm
TL;DR: In this article, two-photon fluorescence excitation (TPE) action is reported for five widely used molecular fluorophores, including Cy 3 (∼140 GM) and Rhodamine 6G ( ∼150GM).
Related Papers (5)
Two-Photon Laser Scanning Fluorescence Microscopy
High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution
Karsten Koenig,I. Riemann +1 more