Square and hexagonal pinholes provide identical image signal levels, if the geometries are compared in a sensible manner. The amount of light passing the pinhole depends on the area of that aperture, consequently the area is the parameter that must be compared when discussing brightness of focus images. The use of incommensurable edge lengths is meant to confuse the reader and thus dishonest and reprehensible. In this article, the signal level as a function of geometry and size in confocal microscopes is described.

Fluorescence recovery after photobleaching (FRAP) has been considered the most widely applied method for observing translational diffusion processes of macromolecules. State of the art laser scanning microscopes such as the Leica TCS SP8 have the advantage of using a high intensity laser for bleaching and a low intensity laser for image recording. The LAS AF application wizard offers different ways to carry out a FRAP experiment.

This webinar will illustrate results obtained by biochemical, Epifluorescence, TIRFM, Confocal and GSD techniques. Depending on the aim of experimental question, different imaging techniques deliver insights into varying aspects of intracellular pathways. To achieve "True-to-detail imaging" of the spatial arrangement of proteins and other biomolecules in cells, GSDIM achieves resolutions up to 20 nm in x and y direction – beyond the diffraction limit of light microscopy. But Super-resolution microscopy can be applied in the axial (z-) direction, too. A recent commercial implementation of the astigmatism approach will be discussed in more detail during this webinar.

This monthly updated reference list demonstrates the major application fields for laser microdissection in life science research.

A deeper understanding of brain function requires visualization of the complex architecture of neurons and their connections. In this special webinar, two researchers working at the cutting-edge of brain imaging will discuss their applications of the latest confocal and two-photon microscopy techniques. From fluorescent labeling and imaging of neural circuits using the Brainbow system to in vivo imaging of brain structure, function, and blood flow, this webinar will give researchers a deeper appreciation of the potential for new microscopy methods to unlock the secrets of the cellular world. Unique insights into sample handling and processing of multicolor images will also be presented, and attendees will have the opportunity to ask questions.

Modern laser microdissection technology has its roots in the early 20th century. It has been steadily advanced and modified over the years. The following article is a summary of the history of laser microdissection from its origins to today’s state of the art.

It has always been a deeply rooted human need to give life a meaning after death. At the end of the 16th century, the friars of the Capuchin monastery in Palermo, Italy, began preserving corpses by embalming them.

This monthly updated references list presents current papers using Leica AM TIRF in the major application fields for TIRF microscopy.

April 26th 2013 is the 170th anniversary of the birth of Ernst Leitz I (1843–1920), who turned a small optical workshop in Wetzlar into an optics company of world renown. At the beginning of the 20th century, the Ernst Leitz Optical Works rapidly became one of the leading suppliers of microscopes and optical products. An exhibition in Wetzlar’s New Town Hall looks back on his life and the milestones of his work. His business skills and his outstanding commitment to his employees and society in general made Ernst Leitz I a sovereign in the best sense of the word: He really was a person who “stood above it all” – not because he disregarded everyone else, but because we simply have to take our hats off to this exceptional personality whose work and actions are far greater than the general notion of a lifetime achievement.

To understand structure and function of brains or other complex biological systems, the method of choice is microscopy. In particular, confocal microscopy is employed to reveal three-dimensional connectivity and functional interactions. To come to a real insight into brain’s way of working, one must look deep into the tissue – which usually is non-transparent. A couple of clearing methods have been developed in the past, but they usually come along with distortions of the structures, incompatibilities with fluorescence stainings or are just prohibitively toxic to the lab technician.

Immersion freezing of thin aqueous specimens is an essential preparation technique for cryo-transmission electron microscopy (cryo-TEM), aiming to preserve fragile biological structures such as molecules and cells in their hydrated environment for a close-to-native visualization. For successful experiments, vitreous ice must be produced, surface contamination must be avoided, and, most important, the natural state of the structure must be preserved.

April 26th 2013 is the 170th anniversary of the birth of Ernst Leitz I (1843-1920), who turned a small optical workshop in Wetzlar into an optics company of world renown. At the beginning of the 20th century, the Ernst Leitz Optical Works rapidly became one of the leading suppliers of microscopes and optical products. This history is illuminated by the two films presented here.

For the understanding of functions of proteins in biological and pathological processes, reporter molecules such as fluorescent proteins have become indispensable tools for visualizing the location of these proteins in intact animals, tissues, and cells. For enzymes, imaging their activity also provides information on their function or functions, which does not necessarily correlate with their location. Metabolic mapping enables imaging of activity of enzymes.

Microscopy-based imaging is booming and the need for tools to retrieve quantitative data from images is urgent. This book provides simple but reliable tools to generate valid quantitative gene expression data, at the mRNA, protein and activity level, from microscopic images in relation to structures in cells, tissues and organs in 2D and 3D.

Especially in fluorescence microscopy, excitation light is friend and foe in one. On the one hand, energy-rich excitation via a specific light wavelength of the fluorochrome resulting in a bright positive fluorochrome signal is highly welcome. On the other hand, "noise" caused by reflections of excitation light passing through the surfaces of optical elements needs to be extremely slight to generate a perfect black background. This relation is described as "signal-to-noise ratio", which is highly relevant for differentiating optically between fluorescence positive and negative cells.