A new type of fluorescent bead is presented by GATTAquant. These beads, called GATTA-Beads, are characterized by a small diameter (23 nm), high intensity and size uniformity. In combination with state-of the-art STED microscopes such as the Leica TCS SP8 STED 3X and high-end image restoration methods available in the Huygens Software, it is shown that these new beads can be used for accurate STED PSF characterization in 3D. Furthermore, it is shown that the measured 3D STED-PSF can be used to improve image restoration quality in combination with STED deconvolution methods available in the Huygens Software.
We have been developing new tools based on fluorogenic forced intercalation (FIT) probes for RNA detection quantification and interference in biological samples. Upon duplex formation with target nucleic acids, the base surrogates TO dye increases its quantum yield and brightness substantially (>10 fold).
Here we show staining of HDAC1 in cancer tissue and epidermoid carcinoma cells. These results clearly show that the use of appropriate validated antibodies and STED microscopy are important tools to study subcellular structures beyond the diffraction limit correcting ill-defined images. This is critical in co-localization studies of proteins inside cells.
At the presynaptic active zone (AZ) a variety of specialized proteins are assembled to complex architectures, which set the basis for speed, precision and plasticity of synaptic transmission. Recently, super-resolution microscopy (SRM) techniques have begun to enter the neurosciences. These approaches combine high spatial resolution with the molecular specificity of fluorescence microscopy. Here, we discuss how SRM techniques can be used to obtain information on the organization of AZ proteins.
The aim of cell biology is to study smallest details on a cellular level preferably in a live cell experiment. By providing fast and direct super-resolution, STED (Stimulated Emission Depletion) microscopy is the perfect tool for studying cellular details in the nanometer range in vivo.
On October 8th 2014, The Royal Swedish Academy of Sciences has decided to award the Nobel Prize in Chemistry for 2014 to Eric Betzig, Stefan W. Hell and William E. Moerner "for the development of super-resolved fluorescence microscopy". For a long time optical microscopy was held back by a presumed limitation: that it would never obtain a better resolution than half the wavelength of light. Helped by fluorescent molecules the Nobel Laureates in Chemistry 2014 ingeniously circumvented this limitation. Their ground-breaking work has brought optical microscopy into the nanodimension.
Professor Stefan Hell is director at the Max Planck Institute for Biophysical Chemistry and head of the department of NanoBiophotonics in Goettingen and widely considered as the father of super-resolution. His inventions of 4Pi and STED microscopy were turned into the first commercial super-resolution microscopes available by Leica Microsystems in 2004 and 2007.
The first super-resolution image he saw was an eye opener for him: "It was not just structures that got smaller. I was looking at a sample that I specifically had high hopes of seeing another layer of complexity and this actually was there."
Stephan Sigrist is professor for biology at the Freie Universität Berlin in Germany. His research focus are synapses, synaptic information transfer and processing between neurons in the developing drosophila larva. His aim is to understand how synapses actually get diversified in our brains.
Yasushi Okada, team leader at Riken Quantitative Biology Center in Osaka, Japan, investigates vesicular transport mechanisms in neuronal cells. As the size of transported vesicles is below 100 nm and the diameter of microtubules is about 25 nm, he uses super-resolution techniques to study the sophisticated machinery of neuronal transport.
Optical imaging devices have a finite depth of field and diffraction limited resolution. The depth of field problem was tackled first with confocal microscopes, diffraction unlimited resolution is available since a few years with super-resolution microscopes. Super-resolution microscopes with a solved depth of field problem are now available.
A research team from Göttingen, led by Prof. Silvio O. Rizzoli, managed to determine the copy numbers and positions of all important building blocks of a synapse for the first time. This allowed them to reconstruct the first scientifically accurate 3D model of a synapse.