Printed in the Netherlands " You have walked to the edge of the great mosaic of human knowledge. Up until now, you have been living in a world full of ideas and concepts that other people have set out for you. Now it is your turn. You get to design a piece of the mosaic and glue it down. It just has to fit with what else is there. And if you do a good job shaping your tile, it will be easier fo...

The spectral resolution of fluorescence microscope images in living cells is achieved by using a confocal laser scanning microscope equipped with grating optics. This capability of temporal and spectral resolution is especially useful for detecting spectral changes of a fluorescent dye; for example, those associated with fluorescence resonance energy transfer (FRET). Using the spectral imaging ...

The human body is a constant flux of thousands of chemical/biological interactions and processes connecting molecules, cells, organs, and fluids, throughout the brain, body, and nervous system. Up until recently it was thought that all these interactions operated in a linear sequence, passing on information much like a runner passing the baton to the next runner. However, the latest findings in...

Fluorescence spectroscopy-based techniques using conventional fluorimeters have been extensively applied since the late 1960s to study different aspects of membrane-related phenomena, i.e., mainly relating to lipid-lipid and lipid-protein (peptide) interactions. Even though fluorescence spectroscopy approaches provide very valuable structurally and dynamically related information on membranes, ...

The fluorescence microscope is one of the most sensitive instruments available for morphological and microchemical analysis of biological material, and especially of cereal grains. Recent innovations in illuminating systems, fluorescence chemistry, and specimen preparation have combined to provide significant improvements over conventional bright-field microscopy in both specificity and sensiti...

We constructed an optical system and developed the software to create and analyze the fluorescence anisotropy images of a variety of objects including single molecules, quantum dots, proteins, nucleic acids and cells. The system includes an inverted microscope, three calcite prisms (CPs) inserted in the excitation and emission optical paths, and CCD camera. The system allows to record dynamic c...

Fluorescence microscopy is an essential tool of modern biology, but, like all forms of optical imaging, it is subject to physical limits on its resolving power. In recent years, several exciting techniques have been introduced to exceed these limits, including standing wave microscopy, 4Pi confocal microscopy, I5M and structured illumination microscopy. Several such techniques have been definit...

Multiphoton fluorescence microscopy has now become a relatively common tool among biophysicists and biologists. The intrinsic sectioning achievable by multiphoton excitation provides a simple means to excite a small volume inside cells and tissues. Multiphoton microscopes have a simplified optical path in the emission side due to the lack of an emission pinhole, which is necessary with normal c...

In fluorescence microscopy, the fluorescence emission can be characterised not only by intensity and position, but also by lifetime, polarization and wavelength. Fluorescence lifetime imaging (FLIM) can report on photophysical events that are difficult or impossible to observe by fluorescence intensity imaging, and time-resolved fluorescence anisotropy imaging (TR-FAIM) can measure the rotation...