Confocal

Pinhole Geometry: Four Corners are Perfect

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.

Confocal Microscopy

May 23, 2013

Tutorial

Step by Step Guide for FRAP Experiments

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.

Quantitative Fluorescence

May 17, 2013

Tutorial

Webinar: Discover the Next Generation of Confocal

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Leica Microsystems recently introduced their next generation of confocal, the Leica TCS SP8 – a truly configurable platform that allows you to create your confocal exactly as you need it. As your scientific research develops further, so you need a system that evolves, adapts and reconfigures. What you discover today will be the foundation of your discoveries tomorrow.

Live-Cell Imaging

Oct 25, 2012

Webinar

Webinar: Challenges in Live Cell Imaging

Live cell imaging is one tool in the cell biologist's tool box. There are, however, a variety of technologies and methods to achieve this. In this webinar, three practicing researchers discuss: Their use of live cell imaging and how it has helped them achieve publishable results.

Live-Cell Imaging

Oct 10, 2012

Webinar

Developments in Multiphoton Excitation Microscopy

Honouring Goeppert-Mayer’s prediction of simultaneous two-photon absorption by an atom or molecule reported in the 1930s in her PhD dissertation thesis, we can state that multiphoton excitation (MPE) microscopy, more frequently identified with two-photon excitation (2PE) fluorescence microscopy, is a key microscopy method in many areas from medicine to biology, from biophysics to materials science.

Multiphoton Microscopy

Oct 08, 2012

Abstract

Webinar: Discover the Next Generation of Confocal – A Platform to Perform to the Highest Level, Whatever Level you Start at

Leica Microsystems recently introduced their next generation of confocal, the Leica TCS SP8 – a truly configurable platform that allows you to create your confocal exactly as you need it. As your scientific research develops further, so you need a system that evolves, adapts and reconfigures. What you discover today will be the foundation of your discoveries tomorrow.

Live-Cell Imaging

Sep 27, 2012

Webinar

Sensors for True Confocal Scanning

In this article, advantages and disadvantages of different types of sensors for single point true confocal scanning devices are discussed. Traditionally, photomultiplier tubes have been employed in such systems. For some cases, avalanche photodiodes have proven to fit best. A new development uniting vacuum and silicon technology has led to chimeric sensors, called hybrid detectors (HyD). They benefit from both technologies.

Confocal Microscopy

Aug 23, 2012

Tutorial

Webinar: Fast Point Scanning Confocal Microscopy

Are you interested in learning more about the latest Leica confocal innovations for high speed imaging and sensitive detection? Then this webinar is for you.

Jun 21, 2012

Webinar

Webinar: Introducing the New Leica TCS SP8 Confocal Platform

Discover Ultra-detailed Imaging with the Leica TCS SP8 ...

Confocal Microscopy

Jun 21, 2012

Webinar

FLCS – Advances in Fluorescence Correlation Spectroscopy

The characterization of substances at the single molecule level has become part of the standard repertoire of scientific research institutes. One of the most common methods is Fluorescence Correlation Spectroscopy (FCS), which can be used to examine the dynamics and concentration of fluorescent molecules in solution.

Confocal Microscopy

Dec 15, 2011

Tutorial

Beam Splitting

Fluorescence Microscopy usually employs incident light illumination. This requires a device that directs the light for illumination into the sample and transmits the light emitted by the sample to the detection system. In the past, various types of mirrors were the only option. Today, the acousto optical beam splitter serves best for the task.

Confocal Microscopy

Dec 09, 2011

Tutorial

Webinar: Frontiers in Confocal Microscopy-Deep Imaging, Vibrational Microscopy (CARS) and High Content Screening Automation

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The free online webinar presented by Leica Microsystems in association with BioOptics World took place on Thursday, December 1 2011. Register and view the webinar on demand.

Dec 01, 2011

Webinar

Neuroscience and Microscopy

Neurobiology, the science of nerves and the brain, has mainly been driven forward in the last 200 years by microscopic investigations. The structures of cellular and subcellular structures, interaction and the three-dimensional assembly of neurons were made visible by various microscopy techniques. The optical microscope is also a necessary tool for visualizing micropipettes in electrophysiological measurements. Thirdly, many types of functional imaging are performed by means of optical microscopy.

Nov 09, 2011

Article

Förster Resonance Energy Transfer (FRET)

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The Förster Resonance Energy Transfer (FRET) phenomenon offers techniques that allow studies of interactions in dimensions below the optical resolution limit. FRET describes the transfer of the energy from an excited state of a donor molecule to an acceptor molecule. Unlike absorption or emission of photons, FRET is a non-radiative energy exchange and consequently not a variation of light-matter interactions.

Confocal Microscopy

Nov 09, 2011

Tutorial

FLIM-FRET in Solutions

FRET efficiency can be measured based on fluorescence lifetime microscopy (FLIM). FLIM-FRET allows analysis of molecular interactions both in vitro and in vivo. This article describes the use of FLIM in the time domain (TCSPC) to measure FRET in vitro in a biochemical assay using a Cerulean-Citrine construct.

Confocal Microscopy

Nov 08, 2011

Article

An Introduction to CARS Microscopy

CARS overcomes the drawbacks of conventional staining methods by the intrinsic characteristics of the method. CARS does not require labeling because it is highly specific to molecular compounds which are based on vibrational contrast and chemical selectivity. The crucial advantage of this method is that the sample remains almost unaffected. With CARS, new samples that could not be stained due to unavailability of the appropriate dye are now accessible.

CARS Microscopy

Nov 04, 2011

Tutorial

Step by Step Guide to Hybrid Detection and Photon Counting

This tutorial explains the underlying hybrid detection technology and compares it to photomultiplier technology. The implications of hybrid detection design for imaging and photon counting are discussed. The tutorial closes with a brief summary of photon counting in the context of imaging.

Live-Cell Imaging

Oct 20, 2011

Tutorial

CARS Publication List

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CARS (Coherent Anti-Stokes Raman Scattering) microscopy is a dye-free method which images structures by displaying the characteristic intrinsic vibrational contrast of their molecules. The crucial advantage of this method is that the sample remains almost unaffected.

CARS Microscopy

Oct 19, 2011

Article

Mosaic Images

Confocal laser scanning microscopes are widely used to create highly resolved 3D images of cells, subcellular structures and even single molecules. Still, an increasing number of scientists are extending their focus of bio-research from single cell studies to entire organs and organisms, analyzing the complex interactions within whole animals.

Confocal Microscopy

Oct 05, 2011

Tutorial

Brighter Fluorescence by Resonant Scanning

Fast True Confocal Scanning reduces photobleaching and increases the fluorescence yield at identical acquisition times. The long-lasting triplet state (or any other “dark state”) is less populated when the illumination is applied in shorter pulses at the same intensity. Consequently, more fluorochromes are available for the fluorescence process (brighter images) and fewer fluorochromes disintegrate from triplet states or excited triplet states (less bleaching).

Confocal Microscopy

Aug 03, 2011

Tutorial

Pinhole Controls Optical Slicing

True confocal scanning microscopes (TCS) use a variable detection pinhole. Good optical sectioning tries to use just the inner core of the PSF.

Confocal Microscopy

Jun 22, 2011

Tutorial

Confocal Optical Section Thickness

Confocal microscopes are employed to optically slice comparably thick samples.

Confocal Microscopy

Jun 21, 2011

Tutorial

Lasers for Confocal

True confocal scanning microscopy (TCS) requires bright diffraction-limited illumination.

Confocal Microscopy

Jun 21, 2011

Tutorial

Webinar: The new Leica HyD

Customize the balance between highest sensitivity and highest dynamic range. The combination of hybrid detection (HyD) with photo-multipliers (PMT) in the same system offers you a dynamic range from single photon counting to imaging whole organisms.

Confocal Microscopy

May 24, 2011

Webinar

Confocal Microscopy

“Confocal Microscopy” refers to a particular optical microscope that allows recording optical sections. Optical sectioning is achieved in a confocal system by illuminating and observing a single diffraction limited spot.

Confocal Microscopy

May 09, 2011

Tutorial

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