In conventional microscopes, fluorescence emission is separated from the backscattered illumination using the Stokes shift, whereby the emission occurs at a longer wavelength to the excitation. Such separation is usually achieved through a combination of wavelength filters that divide the spectrum into mutually exclusive excitation and emission bands. It is therefore impossible in these microsc...

There was an error published in J. Cell Sci. 120, 1703-1705. On p. 1704, in the section ‘Image acquisition’, line 6 should read ‘all four pixels in a 2 2 square are summed into a single pixel’. For clarification, 2 2 binned pixels are twice the height and twice the width of the original pixel, but the pixel area is four times larger; similarly, for a 4 4 binned image the pixel height and width ...

The fluorescence microscope (wide-field, scanning, confocal, one-photon excitation, multiphoton excitation) is an extremely useful and ubiquitous instrument in biological and medical laboratories. The fluorescence microscope provides enhanced contrast, single protein specificity and single molecule sensitivity. Progress in the technical development over more than 100 years of instrument design ...

Bioluminescence microscopy has revealed that gene expression in individual cells can respond differently to the same stimulus. To understand this phenomenon, it is important to sequentially observe the series of events from cellular signal transduction to gene expression regulated by specific transcription factors derived from signaling cascades in individual cells. However, these processes hav...

It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton a...

Most commonly used methods for three-dimensional (3D) fluorescence microscopy make use of sectioning techniques that require that the object be physically scanned in a series of two-dimensional (2D) sections along the z axis. The main drawback in these approaches is the need for these sequential 2D scans. An alternative approach to fluorescence imaging in three dimensions has been developed tha...

We present a new optimization concept for 3D multiphoton fluorescence microscopy by finding the optimal excitation beam giving rise to the smallest possible light-emitting volume or the highest possible signal to noise ratio (SNR).

Dit proefschrift is goedgekeurd door de promotor(en): This work was carried out in graduate school ASCI. ASCI dissertation series number 39. This thesis presents image restoration techniques for applications in (confocal) fluorescence microscopy. We have gained a better understanding of the behavior of non-linear image restoration algorithms and we have developed novel methods to improve their ...

In this research paper, we provide an introduction to fluorescence microscopy techniques. We show how the development of these techniques has lead to significant advances in cellular physiology. As part of this discussion, we detail applications of optical imaging to a study of cell division and to a study of the nervous system. We will conclude by presenting some limitations and controversies ...