Showing papers on "Photoactivated localization microscopy published in 1997"
TL;DR: The present technique provides a simple yet powerful and universal tool for researchers to probe the events of single molecules through objective-type total internal reflection fluorescence microscopy.
334 citations
TL;DR: Three‐dimensional microscopy with nearly isotropic resolution in the λ/5 − λ/10 range is reported, and a comparison with unrestored two‐photon confocal images reveals a total reduction of the uncertainty volume up to a factor of 15.
Abstract: We report three-dimensional (3D) microscopy with nearly isotropic resolution in the lambda/5-lambda/10 range. Our approach combines 4PI-confocal two-photon fluorescence microscopy with image restoration. The 3D resolution is demonstrated with densely clustered beads as well as with F-actin fibers in mouse fibroblast cells. A comparison with unrestored two-photon confocal images reveals a total reduction of the uncertainty volume up to a factor of 15.
142 citations
52 citations
TL;DR: The acquisition of high-quality two-photon fluorescence microscopy images using an all-solid-state self-mode-locked Cr:LiSAF laser is demonstrated and examples of improved depth penetration and reduced dye bleaching are presented.
Abstract: We demonstrate the acquisition of high-quality two-photon fluorescence microscopy images using an all-solid-state self-mode-locked Cr:LiSAF laser. We contrast the performance of the two-photon technique with single-photon confocal fluorescence microscopy images taken with an argon-ion laser. Examples of improved depth penetration and reduced dye bleaching are presented.
17 citations
TL;DR: This overview covers fluorescent molecular probes, filters and filter sets, multiband filters and multidye fluorescence, light sources, microscope objectives, image resolution and the point‐spread function, and general steps for immunolabeling.
Abstract: The growing importance in biology and especially in neurobiology of fluorescence microscopy is due to (1) the extraordinary development of new fluorescent molecular probes and (2) the development of improved low light level imaging systems and confocal microscopy techniques. This overview covers fluorescent molecular probes, filters and filter sets, multiband filters and multidye fluorescence, light sources, microscope objectives, image resolution and the point-spread function, and general steps for immunolabeling.
16 citations
TL;DR: Light microscopy provides a unique tool for examining cell behaviour at the level of the single cell bringing new insight into both cellular heterogeneities and cell-cell interactions.
Abstract: Fluorescence microscopy has become a powerful tool for both the localization of cellular components in fixed cells, using target-specific fluorescent probes and labeled antibodies, and the fluorescence imaging of ions in single living cells. Despite its markedly lower spatial resolution when compared with electron microscopy, the essentially non-invasive nature of light microscopy provides a unique tool for examining cell behaviour at the level of the single cell bringing new insight into both cellular heterogeneities and cell-cell interactions. Further developments in both fluorescent probes and instrumentation should provide even more powerful tools to probe the mechanisms of cell function, both in vitro and in vivo.
8 citations
Dissertation•
02 Dec 1997
7 citations
5 citations
10 Apr 1997
TL;DR: In this article, a simple approach to ameliorating the effects of bleaching is presented, which is to progressively increase the integration times so as to maintain constant signal level from slice to slice.
Abstract: Photobleaching causes progressive fading in successive slices of a through-focus series, so that the last image taken in the series can have a significantly lower signal-to-noise ratio than the first. Bleaching is oftensuccessfully minimized by including antifade additives in the specimen preparation and/or by reducing theoptical dose to the specimen. However, these measures may not be sufficient in a through-focus series where many slices must be taken. This paper presents a simple approach to ameliorating the effects of bleaching, which is to progressively increase the integration times so as to maintain constant signal level from slice to slice. I refer to the sequence of integration times as an integration schedule. I develop the equations for integration scheduling from the physical assumptions and discuss how the method affects image quality.Keywords: microscopy, fluorescence, photobleaching, confocal, three-dimensional microscopy, photodetector 1. INTRODUCTION In fluorescence microscopy, a structure or function of interest in a specimen is labeled with a fluorescent probe.
29 Dec 1997
TL;DR: The last 20 years have seen an unexpected great renaissance and a partial revolution in light microscopy, due to new design in optics and instrumentation as well as improvement of optical contrast enhancement techniques.
Abstract: The last 20 years have seen an unexpected great renaissance and a partial revolution in light microscopy. This recent progress is due to new design in optics and instrumentation as well as improvement of optical contrast enhancement techniques. Recent progress in fluorescence microscopy is achieved by multiparameter fluorescence techniques, by improvement of conventional photomicrography as well as by optoelectronic imaging, confocal laser scanning microscopy, image processing and analysis. Due to the increase in number of fluorescence dyes, double and triple bandpass filter sets permit a rapid changeover between different fluorochromes simultaneously.© (1997) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.