Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Microscopy: Nine at one blow

04.02.2016

Advance in biomedical imaging: The University of Würzburg's Biocenter has enhanced fluorescence microscopy to label and visualise up to nine different cell structures simultaneously.

Fluorescence microscopy allows researchers to visualise biomolecules in cells. They label the molecules using fluorescent probes, excite them with light and use the fluorescence thus triggered to get a microscopic image of the cell's structures.


The sFLIM procedure enables labelling three different cell structures at once using the same fluorescent dye, allowing them to be distinguished clearly.

(Picture: Thomas Niehörster)


Unique: Nine different cell structures were fluorescence labelled at once to become distinguishable under the microscope.

(Picture: Thomas Niehörster)

"A major difficulty in this process is to clearly distinguish the many fluorescent probes that are often quite similar to one another," says Thomas Niehörster, a doctoral student of Professor Markus Sauer at the Department of Biotechnology and Biophysics of the Julius-Maximilians-Universität Würzburg (JMU) in Bavaria, Germany.

What the new strategy is based on

To make progress in this field, the scientists have devised a new strategy. For excitation of the probes, they now use three alternately pulsed lasers of different wavelengths (blue, green and red). Additionally, they benefit from differences in the probes' emission spectrum and the slightly different fluorescence decay patterns over time in the range of a few nanoseconds.

This test setup delivers complex data which the scientists analyse using their own software. The entire process is called sFLIM (spectrally resolved fluorescence lifetime microscopy) and seems to work flawlessly: "It enables us to distinguish the fluorescent probes with unparalleled accuracy," Professor Sauer says.

What the method is capable of

This approach allows five fluorescent probes to be distinguished with each of the three lasers, resulting in 15 different structures that could theoretically be visualised at the same time.

In practice, however, the scientists have to settle for less: "It is difficult to label so many different cell structures at once, and there is only a limited number of probes to do so," Niehörster explains. "Nevertheless, we managed to label and visualise nine different structures simultaneously." This includes, for instance, the F-Actin protein structure of the cytoskeleton, the nuclear membrane or newly created DNA.

The method's high sensitivity moreover allows using the same fluorescent dye to label three different cell structures at once that can be distinguished clearly afterwards. This is because fluorescence properties vary slightly depending on the chemical environment in the cell and become distinguishable as a result.

Cooperation with science and industry

This result was accomplished through a partnership of the JMU Department with the University of Göttingen (software) and the company PicoQuant in Berlin (hardware architecture). It was published in the scientific journal "Nature Methods".

"Multi-target spectrally resolved fluorescence lifetime imaging microscopy", Thomas Niehörster, Anna Löschberger, Ingo Gregor, Benedikt Krämer, Hans-Jürgen Rahn, Matthias Patting, Felix Koberling, Jörg Enderlein & Markus Sauer, Nature Methods (2016), doi:10.1038/nmeth.3740

Contact

Prof. Dr. Markus Sauer, Department of Biotechnology and Biophysics, University of Würzburg, Phone +49 931 31-88687, m.sauer@uni-wuerzburg.de

Robert Emmerich | Julius-Maximilians-Universität Würzburg
Further information:
http://www.uni-wuerzburg.de

More articles from Life Sciences:

nachricht Ion treatments for cardiac arrhythmia — Non-invasive alternative to catheter-based surgery
20.01.2017 | GSI Helmholtzzentrum für Schwerionenforschung GmbH

nachricht Seeking structure with metagenome sequences
20.01.2017 | DOE/Joint Genome Institute

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Traffic jam in empty space

New success for Konstanz physicists in studying the quantum vacuum

An important step towards a completely new experimental access to quantum physics has been made at University of Konstanz. The team of scientists headed by...

Im Focus: How gut bacteria can make us ill

HZI researchers decipher infection mechanisms of Yersinia and immune responses of the host

Yersiniae cause severe intestinal infections. Studies using Yersinia pseudotuberculosis as a model organism aim to elucidate the infection mechanisms of these...

Im Focus: Interfacial Superconductivity: Magnetic and superconducting order revealed simultaneously

Researchers from the University of Hamburg in Germany, in collaboration with colleagues from the University of Aarhus in Denmark, have synthesized a new superconducting material by growing a few layers of an antiferromagnetic transition-metal chalcogenide on a bismuth-based topological insulator, both being non-superconducting materials.

While superconductivity and magnetism are generally believed to be mutually exclusive, surprisingly, in this new material, superconducting correlations...

Im Focus: Studying fundamental particles in materials

Laser-driving of semimetals allows creating novel quasiparticle states within condensed matter systems and switching between different states on ultrafast time scales

Studying properties of fundamental particles in condensed matter systems is a promising approach to quantum field theory. Quasiparticles offer the opportunity...

Im Focus: Designing Architecture with Solar Building Envelopes

Among the general public, solar thermal energy is currently associated with dark blue, rectangular collectors on building roofs. Technologies are needed for aesthetically high quality architecture which offer the architect more room for manoeuvre when it comes to low- and plus-energy buildings. With the “ArKol” project, researchers at Fraunhofer ISE together with partners are currently developing two façade collectors for solar thermal energy generation, which permit a high degree of design flexibility: a strip collector for opaque façade sections and a solar thermal blind for transparent sections. The current state of the two developments will be presented at the BAU 2017 trade fair.

As part of the “ArKol – development of architecturally highly integrated façade collectors with heat pipes” project, Fraunhofer ISE together with its partners...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

Sustainable Water use in Agriculture in Eastern Europe and Central Asia

19.01.2017 | Event News

12V, 48V, high-voltage – trends in E/E automotive architecture

10.01.2017 | Event News

2nd Conference on Non-Textual Information on 10 and 11 May 2017 in Hannover

09.01.2017 | Event News

 
Latest News

Helmholtz International Fellow Award for Sarah Amalia Teichmann

20.01.2017 | Awards Funding

An innovative high-performance material: biofibers made from green lacewing silk

20.01.2017 | Materials Sciences

Ion treatments for cardiac arrhythmia — Non-invasive alternative to catheter-based surgery

20.01.2017 | Life Sciences

VideoLinks
B2B-VideoLinks
More VideoLinks >>>