Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

From Microscopy to Nanoscopy

10.08.2007
Photoswitchable rhodamine amides for high-resolution optical 3D far-field microscopy

Layer-by-layer light microscopic nanoscale images of cells and without having to prepare thin sections?

A team led by Stefan Hell and Mariano Bossi at the Max Planck Intstitute for Biophysical Chemistry in Göttingen is now leading the way with a technique called optical 3D far-field microscopy—with nanoscale resolution, good signal-to-noise ratio, and relatively short exposure times.

The secret of their success lies in special photoswitchable fluorescence dyes, the researchers report to the journal Angewandte Chemie. These rhodamine amides make it possible to obtain highly resolved 3D images of transparent fluorescence-marked samples such as living cells.

... more about:
»Photon »amides »rhodamine »switched »wavelength

Until fairly recently, the resolution of light microscopes was limited by the wavelength of the light. This means that details finer than 200 nanometers (millionths of millimeters) cannot be observed. There are non-optical methods, such as electron microscopy, but light microscopy is still the only way to observe the interior of whole, or even living, cells. The use of fluorescent dyes makes it possible to selectively obtain images of individual cell components, for example, proteins. Today, the wavelength dogma is overcome.

Hell received the German Future Prize in 2006 for the first concept breaking the wavelength barrier the stimulated emission depletion (STED) microscope . Molecules are transferred from a “dark” (non-fluorescent) to a “bright” (fluorescent) excited energy state—with a spacial sharpness far beyond those 200 nanometers.

Now the german team is demonstrating the power of another concept. They use molecules that are not only transferred but can be “switched” from “fluorescent” to “non-fluorescent” and back. In contrast to the STED and other related methods of the team, only separate, isolated marker molecules are randomly switched on at the same time. Their fluorescence is registered, and then they get switched off again automatically. In this way, the simultaneously fluorescing (switched on) markers are farther apart from each other than the minimum distance that the microscope can resolve.

This is only possible using switchable molecules that emit many photons, one after the other, when switched on. If these photons are captured with a camera, the centers of the individual fluorescing dots can be distinguished. After the exposure, the molecule becomes dark again (switches off), allowing further, neighboring molecules to be photographed. This process is repeated many times, until many dots become a picture. The full distribution can be reconstructed—at a resolution not limited by the wavelength of light.

The researchers have now found a class of substances that fulfill all the requirements of this technique: rhodamine amides. At the core of these molecules lies a system of five rings. In this form, the compound is colorless and does not fluoresce. Irradiation with light induces an isomerization in which one of the rings is opened. This form of the molecule is red and can be excited several times.

Most importantly: rhodamine amides can be switched on by either a UV photon or two photons in the red part of the spectrum. This two-photon excitation can be focused onto a thin plane, which allows biological samples to be photographed layer by layer. The individual images can then be reconstructed into a single multilayer image. The resolution reached in the focal plane is far beyond the diffraction barrier (10–30 nm).

Author: Stefan W. Hell, Max-Planck-Institut für Biophysikalische Chemie, Göttingen (Germany), http://www.mpibpc.gwdg.de/abteilungen/200/

Title: Photochromic Rhodamines Provide Nanoscopy with Optical Sectioning

Angewandte Chemie International Edition 2007, 46, No. 33, 6266–6270, doi: 10.1002/anie.200702167

Stefan W. Hell | Angewandte Chemie
Further information:
http://www.mpibpc.gwdg.de/abteilungen/200/
http://pressroom.angewandte.org

Further reports about: Photon amides rhodamine switched wavelength

More articles from Life Sciences:

nachricht Modern genetic sequencing tools give clearer picture of how corals are related
17.08.2017 | University of Washington

nachricht The irresistible fragrance of dying vinegar flies
16.08.2017 | Max-Planck-Institut für chemische Ökologie

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: Fizzy soda water could be key to clean manufacture of flat wonder material: Graphene

Whether you call it effervescent, fizzy, or sparkling, carbonated water is making a comeback as a beverage. Aside from quenching thirst, researchers at the University of Illinois at Urbana-Champaign have discovered a new use for these "bubbly" concoctions that will have major impact on the manufacturer of the world's thinnest, flattest, and one most useful materials -- graphene.

As graphene's popularity grows as an advanced "wonder" material, the speed and quality at which it can be manufactured will be paramount. With that in mind,...

Im Focus: Exotic quantum states made from light: Physicists create optical “wells” for a super-photon

Physicists at the University of Bonn have managed to create optical hollows and more complex patterns into which the light of a Bose-Einstein condensate flows. The creation of such highly low-loss structures for light is a prerequisite for complex light circuits, such as for quantum information processing for a new generation of computers. The researchers are now presenting their results in the journal Nature Photonics.

Light particles (photons) occur as tiny, indivisible portions. Many thousands of these light portions can be merged to form a single super-photon if they are...

Im Focus: Circular RNA linked to brain function

For the first time, scientists have shown that circular RNA is linked to brain function. When a RNA molecule called Cdr1as was deleted from the genome of mice, the animals had problems filtering out unnecessary information – like patients suffering from neuropsychiatric disorders.

While hundreds of circular RNAs (circRNAs) are abundant in mammalian brains, one big question has remained unanswered: What are they actually good for? In the...

Im Focus: RAVAN CubeSat measures Earth's outgoing energy

An experimental small satellite has successfully collected and delivered data on a key measurement for predicting changes in Earth's climate.

The Radiometer Assessment using Vertically Aligned Nanotubes (RAVAN) CubeSat was launched into low-Earth orbit on Nov. 11, 2016, in order to test new...

Im Focus: Scientists shine new light on the “other high temperature superconductor”

A study led by scientists of the Max Planck Institute for the Structure and Dynamics of Matter (MPSD) at the Center for Free-Electron Laser Science in Hamburg presents evidence of the coexistence of superconductivity and “charge-density-waves” in compounds of the poorly-studied family of bismuthates. This observation opens up new perspectives for a deeper understanding of the phenomenon of high-temperature superconductivity, a topic which is at the core of condensed matter research since more than 30 years. The paper by Nicoletti et al has been published in the PNAS.

Since the beginning of the 20th century, superconductivity had been observed in some metals at temperatures only a few degrees above the absolute zero (minus...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

Call for Papers – ICNFT 2018, 5th International Conference on New Forming Technology

16.08.2017 | Event News

Sustainability is the business model of tomorrow

04.08.2017 | Event News

Clash of Realities 2017: Registration now open. International Conference at TH Köln

26.07.2017 | Event News

 
Latest News

Gold shines through properties of nano biosensors

17.08.2017 | Physics and Astronomy

Greenland ice flow likely to speed up: New data assert glaciers move over sediment, which gets more slippery as it gets wetter

17.08.2017 | Earth Sciences

Mars 2020 mission to use smart methods to seek signs of past life

17.08.2017 | Physics and Astronomy

VideoLinks
B2B-VideoLinks
More VideoLinks >>>