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Department of Chemical Engineering and Biotechnology

Q&A Session: 14:15 - 14:45 on Wednesday 23rd September

Fluorescent Microscopy and Stochastic Optical Reconstruction Microscopy (STORM)

What Will Students Will See in the Video Demo?

In Biotechnology physics plays a major role as many physics techniques help characterise and analyse biological objects (for example cell, bacteria etc.). One of the main physics techniques used in studying biological samples are different kind of microscopes. This video demo explains the principles of physics working behind fluorescent microscopy, the most common microscopy technique used in science in order to observe biological objects. At the Department of Chemical Engineering and Biotechnology Laser Analytics and Molecular Neuroscience lab groups there is also a special super-resolution fluorescent microscopy technique being used: Stochastic Optical Reconstruction Microscopy (STORM).

What Physics is Used?

Fluorescent microscopy is based on ‘fluorescence’ – the ability of the substance to emit light due to its light absorption properties. By attaching fluorescent molecules (or fluorophores) to specific cell organelles or other structures one can uniquely observe the structures of interest. All optical microscopes (including standard fluorescent microscope) have a resolution limit defined by Ernst Abbe. This limit, however, can be overcome in a number of super-resolution fluorescent microscopy techniques. STORM is one of such techniques, where the resolution of 20 nm could be achieved. In STORM fluorescent molecules are blinking instead of emitting continuous light. This is needed in order to decrease the number of shining fluorophores in one frame, which will allow to pinpoint the precise location of each molecule. Then, in order to reconstruct the image, one needs to record a big number of frames (tens of thousands), as at one frame only some small portion of all fluorescent molecules will emit light. Reconstructed image is then produced by adding up blinks from all recorded frames.

Why is this Useful?

With the cutting-edge super-resolution technique (STORM) this microscopy helps locate specific cellular components, organelles, proteins etc… with great precision. In our Laser lab this technique is used to study the location of proteins inside neural synapse, which is only about one micron (one millionth of a meter) in size!



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