Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:


From Microscopy to Nanoscopy

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),

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:

Further reports about: Photon amides rhodamine switched wavelength

More articles from Life Sciences:

nachricht First time-lapse footage of cell activity during limb regeneration
25.10.2016 | eLife

nachricht Phenotype at the push of a button
25.10.2016 | Institut für Pflanzenbiochemie

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Etching Microstructures with Lasers

Ultrafast lasers have introduced new possibilities in engraving ultrafine structures, and scientists are now also investigating how to use them to etch microstructures into thin glass. There are possible applications in analytics (lab on a chip) and especially in electronics and the consumer sector, where great interest has been shown.

This new method was born of a surprising phenomenon: irradiating glass in a particular way with an ultrafast laser has the effect of making the glass up to a...

Im Focus: Light-driven atomic rotations excite magnetic waves

Terahertz excitation of selected crystal vibrations leads to an effective magnetic field that drives coherent spin motion

Controlling functional properties by light is one of the grand goals in modern condensed matter physics and materials science. A new study now demonstrates how...

Im Focus: New 3-D wiring technique brings scalable quantum computers closer to reality

Researchers from the Institute for Quantum Computing (IQC) at the University of Waterloo led the development of a new extensible wiring technique capable of controlling superconducting quantum bits, representing a significant step towards to the realization of a scalable quantum computer.

"The quantum socket is a wiring method that uses three-dimensional wires based on spring-loaded pins to address individual qubits," said Jeremy Béjanin, a PhD...

Im Focus: Scientists develop a semiconductor nanocomposite material that moves in response to light

In a paper in Scientific Reports, a research team at Worcester Polytechnic Institute describes a novel light-activated phenomenon that could become the basis for applications as diverse as microscopic robotic grippers and more efficient solar cells.

A research team at Worcester Polytechnic Institute (WPI) has developed a revolutionary, light-activated semiconductor nanocomposite material that can be used...

Im Focus: Diamonds aren't forever: Sandia, Harvard team create first quantum computer bridge

By forcefully embedding two silicon atoms in a diamond matrix, Sandia researchers have demonstrated for the first time on a single chip all the components needed to create a quantum bridge to link quantum computers together.

"People have already built small quantum computers," says Sandia researcher Ryan Camacho. "Maybe the first useful one won't be a single giant quantum computer...

All Focus news of the innovation-report >>>



Event News

#IC2S2: When Social Science meets Computer Science - GESIS will host the IC2S2 conference 2017

14.10.2016 | Event News

Agricultural Trade Developments and Potentials in Central Asia and the South Caucasus

14.10.2016 | Event News

World Health Summit – Day Three: A Call to Action

12.10.2016 | Event News

Latest News

Greater Range and Longer Lifetime

26.10.2016 | Power and Electrical Engineering

VDI presents International Bionic Award of the Schauenburg Foundation

26.10.2016 | Awards Funding

3-D-printed magnets

26.10.2016 | Power and Electrical Engineering

More VideoLinks >>>