Sep. 25, 2018

Read & Win: Fluorescence Microscopy - From Principles to Biological Applications

  • Fluorescence Microscopy - From Principles to Biological ApplicationsFluorescence Microscopy - From Principles to Biological Applications
  • Fluorescence Microscopy - From Principles to Biological Applications
  • Prof. Dr. Ulrich Kubitschek

Fluorescence Microscopy has become a very powerful tool that allows to specifically visualize structure or proteins in cells. During the last 20 years, many novel techniques have emerged to enhance spatial and or temporal resolution of live fluorescence probes. These techniques now allow for the detection of single molecules and a spatial resolution which is below the theoretical resolution limit for traditional light microscopy, approaching the resolution of electron microscopy.

This book, now in its second edition, is specifically designed to allow advanced students and researchers with little background in physics to comprehend both microscopy basics and state-of the-art light microscopy techniques. Each chapter is written by a renowned expert in the field, and has been thoroughly revised to include more didactical features: Theoretical derivations have been concentrated in text boxes and summaries provide fast access for beginners. The supplementary material has been expanded, with a companion website that offers additional teaching material including videos.

Win the book!
To have a chance of winning the book read Issue 3, 2018 of Imaging & Microscopy (page 12). As a subscriber you could read the issue already online or order you own copy (as a free trial copy). Take part in our competition and send your answer to with the subject line Read & Win. All correct answers will be entered in a prize draw and the lucky winner will receive a copy of "Fluorescence Microscopy - From Principles to Biological Applications".
Closing date: November 7, 2018.

Prof. Dr. Ulrich Kubitscheck
Ulrich Kubitscheck studied physics at the University of Bremen, Germany, and the University of Maryland at College Park, USA, before his postdoctoral trainings at The Weizmann Institute of Science, Israel, and the University of Münster, Germany. He took up his present appointment as professor for physical chemistry at the University of Bonn in 2004.

During his career Ulrich Kubitscheck applied numerous of the major light microscopy techniques. He has authored over 80 scientific publications and has extensive experience in teaching courses on physical chemistry, biophysics and quantitative microscopy. Currently, he focuses on RNA export, and develops single molecule and light sheet fluorescence imaging techniques.

Interview with Ulrich Kubitschek:

What is your main focus in research, what is your main scientific interest?

Kubitscheck: It fascinates me to observe fundamental cellular processes and structures at utmost resolution. On the one hand this means that I am thrilled by elucidating molecular processes like RNA export at the single molecule level. On the other hand, this means currently for me to visualize neuronal structures and interconnections over large distances at highest possible optical resolution. We achieve this goal by combining light sheet fluorescence microscopy to achieve a high imaging speed over large distances, expansion microscopy to realize a high effective resolution, and genetic labeling tools to distinguish synaptically connected neurons. Advanced light microscopy always plays a major role in the science I am doing.

What was the reason to edit the second edition of this book?

Kubitscheck: The first edition still missed several relevant microscopy approaches: two-photon excitation, light sheet fluorescence microscopy and localization microscopy. For the second edition, I could convince experts in each of these fields to join the author board and received excellent contributions. Furthermore, some of the chapters of the first edition were not as accessible as intended. This was reflected by reader feedbacks. The respective authors now invested a lot of time and effort to clarify these. Of course, all chapters were revised for clarity and novelty. I am really very pleased by the final product. This is the book I intended to make!

What is the target audience for the book?

Kubitscheck: The people that use advanced light microscopy for their research: students, postdocs or researchers in the life sciences. No current curriculum covers the material that is presented in the book. Whoever uses one of the modern light microscopy techniques needs a basic understanding how they work in order to evaluate their strengths, weaknesses and pitfalls.

What knowledge is prerequisite for the book?

Kubitscheck: For the major part a very basic knowledge of physics is sufficient. To make the material intuitively accessible is one of the main intentions of the book. The most important basics of optics are repeated in chapter 1. However, to really understand complex techniques like structural illumination microscopy or fluorescence resonance energy transfer a profound math knowledge is required. But for most of the book some physical intuition and the desire to understand is completely sufficient.

What is the structure of the book?

Kubitscheck: The structure was completely revised in the new edition. We start out with the basics: optics, classical light microscopy, fluorescence microscopy and finally labeling techniques. Then we continue with the optical sectioning techniques, confocal, two photon and light sheet microscopy, followed by the super resolution techniques: localization-based, SIM and STED microscopy. Finally, the advanced quantitative techniques are discussed: photobleaching, single molecule microscopy and FRET. The book contains also very useful appendices on the basics of image processing and practical optical alignment. The latter reports quite a number of very useful tricks for people that develop their own instruments.

Which methods have developed the most since the last edition of your book?

Kubitscheck: Light sheet fluorescence microscopy really took off in the past years. Several different configurations became recently commercially available, with more to come. The new detectors being used in the last generation of confocal microscopes are very hot. Furthermore, two photon and fluorescence life time imaging are really spreading out and became accessible for non-experts. Big data, image storage and data transfer are becoming topics of increasing importance. Too new even for the second edition was Stefan Hell´s second stroke, MINFLUX.

If you could advance one particular technique, which would it be?

Kubitscheck: I am particularly fascinated by the options of lattice light sheet microscopy. It offers a number of great advantages, yet is obviously not easy to master.

How important is automated image analysis?

Kubitscheck: Quantitative research using microscopy is based on automated image analysis. Image data became so huge that it cannot be handled anymore just manually. At my institute we constantly invest in improving the image processing hard- and software. Automated image analysis using FIJI, Mathlab and further commercial products play a huge role. Unfortunately, the race between the available capabilities and the requirements appears to me as the race between the hare and the hedgehog ...

Fluorescence Microscopy
From Principles to Biological Applications

Ulrich Kubitscheck (Editor)

2nd Edition, June 2017
ISBN: 978-3-527-33837-5
Wiley-VCH, Weinheim
Also available in digital formats!
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