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 Newly discovered bacteria-binding protein in the intestine
08.12.2016 | University of Gothenburg

nachricht The balancing act: An enzyme that links endocytosis to membrane recycling
07.12.2016 | National Centre for Biological Sciences

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Significantly more productivity in USP lasers

In recent years, lasers with ultrashort pulses (USP) down to the femtosecond range have become established on an industrial scale. They could advance some applications with the much-lauded “cold ablation” – if that meant they would then achieve more throughput. A new generation of process engineering that will address this issue in particular will be discussed at the “4th UKP Workshop – Ultrafast Laser Technology” in April 2017.

Even back in the 1990s, scientists were comparing materials processing with nanosecond, picosecond and femtosesecond pulses. The result was surprising:...

Im Focus: Shape matters when light meets atom

Mapping the interaction of a single atom with a single photon may inform design of quantum devices

Have you ever wondered how you see the world? Vision is about photons of light, which are packets of energy, interacting with the atoms or molecules in what...

Im Focus: Novel silicon etching technique crafts 3-D gradient refractive index micro-optics

A multi-institutional research collaboration has created a novel approach for fabricating three-dimensional micro-optics through the shape-defined formation of porous silicon (PSi), with broad impacts in integrated optoelectronics, imaging, and photovoltaics.

Working with colleagues at Stanford and The Dow Chemical Company, researchers at the University of Illinois at Urbana-Champaign fabricated 3-D birefringent...

Im Focus: Quantum Particles Form Droplets

In experiments with magnetic atoms conducted at extremely low temperatures, scientists have demonstrated a unique phase of matter: The atoms form a new type of quantum liquid or quantum droplet state. These so called quantum droplets may preserve their form in absence of external confinement because of quantum effects. The joint team of experimental physicists from Innsbruck and theoretical physicists from Hannover report on their findings in the journal Physical Review X.

“Our Quantum droplets are in the gas phase but they still drop like a rock,” explains experimental physicist Francesca Ferlaino when talking about the...

Im Focus: MADMAX: Max Planck Institute for Physics takes up axion research

The Max Planck Institute for Physics (MPP) is opening up a new research field. A workshop from November 21 - 22, 2016 will mark the start of activities for an innovative axion experiment. Axions are still only purely hypothetical particles. Their detection could solve two fundamental problems in particle physics: What dark matter consists of and why it has not yet been possible to directly observe a CP violation for the strong interaction.

The “MADMAX” project is the MPP’s commitment to axion research. Axions are so far only a theoretical prediction and are difficult to detect: on the one hand,...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

ICTM Conference 2017: Production technology for turbomachine manufacturing of the future

16.11.2016 | Event News

Innovation Day Laser Technology – Laser Additive Manufacturing

01.11.2016 | Event News

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

14.10.2016 | Event News

 
Latest News

Scientists track chemical and structural evolution of catalytic nanoparticles in 3-D

08.12.2016 | Materials Sciences

Decoding cement's shape promises greener concrete

08.12.2016 | Materials Sciences

Will Earth still exist 5 billion years from now?

08.12.2016 | Physics and Astronomy

VideoLinks
B2B-VideoLinks
More VideoLinks >>>