Advanced Functional Imaging Workshop

30. 5. 2018, 10:45 - 14:30
room 211, building A35, CEITEC MU, University Campus Bohunice

Are you having an interesting biological problem and need to demonstrate and/or functionally characterize the studied phenomenon in living cells?

Are you working with plant or human/animal systems?

Do you do the curiosity-driven fundamental or applied research and development?

If you have answered at least one of the aforementioned questions YES, then you are a good candidate for the participation of our brief (3 hours) workshop.

 

 

The main workshop goals are:

  • Providing you with the recent advances in optical microscopy techniques for functional imaging, available both in Vienna and in Brno via services/collaboration provided by VBCF/CEITEC, respectively
  • Highlighting practical aspects, discussion of your problems/needs will be highly welcome

REGISTER BELOW.

Program

10:45 Jan Hejátko: Welcome
11:00 – 12:00 Kareem Elsayad: Beyond Conventional Optical Microscopy
12:00 – 12:30 Lunch/Refreshments
12:30 – 13:30 Aswathy Jayasree: FLIM-Based Functional Imaging
13:30 Visit of CELLIM CF/Demonstration of FLIM Microscope (Zeiss LSM 780 with Becker&Hickl FLIM/FCS and “white” In Tune pulse laser)

 

 

Kareem Elsayad, head of the Advanced Microscopy CF at VBCF, Vienna, Austria

Beyond Conventional Optical Microscopy

Here I will give an overview of some cutting edge and novel optical microscopy techniques beyond those that can be found in a microscopy core facility and which are available at The VBCF Advanced Microscopy Facility. In particular, I will focus on the use of time- and wavelength- resolved methods to extract information on the dynamics, structure and mechanical properties of biological samples (1), as well as some novel illumination and detection schemes that can be used for higher resolution/contrast time-lapse imaging.


 

Aswathy Jayasree, post-doc and advanced imaging specialist in the Functional Genomics and Proteomics of Plants group, CEITEC MU, Brno, Czech Republic

FLIM-Based Functional Imaging

Fluorescence Lifetime Imaging (FLIM) is recording the fluorescence lifetime of a molecule as an image instead of photon intensity. Since fluorescence lifetime is characteristic of a particular fluorophore, this information can be used to distinguish between two molecules that emit the fluorescence of the same wavelength and their immediate (functional) status. 
FLIM-based imaging will be demonstrated for i) quantification and localization of protein-protein interactions in vivo at subcellular resolution ii) functional imaging using specific reporters for e.g. macromolecular crowding (2) and iii) Fluorescence Intensity Decay Shape Analysis Microscopy (FIDSAM) to filter out a strong autofluorescence background from the real fluorescent signal which is very difficult to distinguish using a regular confocal or epifluorescent microscope. This method is an alternative particularly in plant systems to spectral unmixing frequently used in animals that allows subtracting the specific signal even if the signal intensity reaches about 10% of the noise (3). 


 

References

  1. Elsayad, K., et al. (2016). Sci Signal 9, rs5.
  2. Boersma, A. J., et al. (2015). Nat Methods 12, 227-229, 221 p following 229.
  3. Schleifenbaum, F., et al. (2010). Mol Plant 3, 555-562.