3-DIMENSIONAL SUPERRESOLUTION WITH A 4PI-CONFOCAL MICROSCOPE USING IMAGE-RESTORATION

Citation
M. Schrader et al., 3-DIMENSIONAL SUPERRESOLUTION WITH A 4PI-CONFOCAL MICROSCOPE USING IMAGE-RESTORATION, Journal of applied physics, 84(8), 1998, pp. 4033-4042
Citations number
43
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Physics, Applied
Journal title
ISSN journal
0021-8979
Volume
84
Issue
8
Year of publication
1998
Pages
4033 - 4042
Database
ISI
SICI code
0021-8979(1998)84:8<4033:3SWA4M>2.0.ZU;2-N
Abstract
The combination of two-photon excitation 4Pi-confocal fluorescence mic roscopy with image restoration leads to a fundamental improvement of t hree-dimensional resolution in the imaging of transparent, fluorescent specimens. The improvement is exemplified by randomly dispersed fluor escent beads and with actin filaments in a mouse fibroblast cell. For an illumination wavelength of 810 nm, we obtained lateral and axial fu ll-width at half-maxima of point-like objects of 120-140 nm, and 70-10 0 nm, respectively. Fluorescent beads that are 150 nm apart are imaged with an intensity dip of similar to 25%. This amounts to a improvemen t of the axial resolution over;sixfold standard two-photon confocal mi croscopy. In the cell, the 3D-images reveal details otherwise not reso lvable with focused light. (C) 1998 American Institute of Physics. [S0 021-8979(98)00920-7].