Laboratory of Advanced Optical Microscopy
Welcome & What can we offer
The Laboratory of Advanced Optical Microscopy (LAOM), affiliated with the Center for Systems Biology (C4Sys), provides facilities for advanced optical microscopy experiments. Our specialty is two-photon polarization microscopy (2PPM), a technique we have developed and continue developing.
The technique (also known as fluorescence-detected linear dichroism) is described in detail in our Nature Methods publication, freely available at http://www.nh.cas.cz/people/lazar/celler/downloads.php. A less technical explanation is available on our webpages (http://www.nh.cas.cz/people/lazar/celler/absorption.php), as well as on the website of Innovative Bioimaging, L.L.C. (www.innovativebioimaging.com).
2PPM is a powerful microscopy technique suitable for observing membrane proteins and cytoskeletal proteins labeled with a fluorescent moiety (such as a fluorescent protein). Two-photon polarization microscopy detects the angle at which the fluorescent moiety is oriented with respect to the cell membrane (or cytoskeleton). If the mean tilt angle (or the distribution of the tilt angle) changes during a molecular process, you can observe this process by using two-photon polarization microscopy.
For example, the technique has been shown to be exceptionally sensitive in direct detection of G-protein activation, since the activation-induced changes in interactions between Gα and Gβγ subunits generally affect the orientational freedom of a fluorescent label attached to the Gα subunit increases. In contrast to other fluorescence microscopy techniques (namely FRET), 2PPM only needs a single fluorescent label in order to detect protein-protein interactions and their changes.
The laboratory is equipped by a highly customized two-photon laser scanning polarization microscope based on an iMic system by Till Photonics GmbH/FEI GmbH. The setup also allows single-photon confocal polarization microscopy observatitions. It includes a wide range of laser light sources (Chameleon Ultra II, 405nm, 488nm, 534nm, 632nm), as well as equipment for wide field fluorescence observation. Furthermore, the laboratory has access to a single- and two-photon confocal polarization microscope based on the Olympus Fluoview IX81 platform. The laboratory is capable of performing a variety of combined technique experiments that involve microscopy along with physiology, pharmacology or electrophysiology.
The Laboratory of Advanced Optical Microscopy is located within the Czech Academy of Sciences campus in Nove Hrady, Czech Republic, in a beautiful chateau that houses the Center of Nanobiology and Structural Biology of the Microbiology Institute CAS.
How can you access
General questions
Josef Lazar, lazar@nh.cas.cz,
phone: +420 723415295
Head: Josef Lazar (lazar@nh.cas.cz)
Microscopy specialists: Alexey Bondar, Ievgeniia Iermak
Staff: Karolína Tošnerová, Olga Rybakova
Customer registration
Please, fill in the short registration form
The information will be used (i) for your identification as a customer of our infrastructure (ii) for contacting you in case we will need more information about your samples.
Services
The laboratory provides imaging services performed using living or fixed mammalian cells, yeast, and multicellular organisms (C. elegans, D. melanogaster and others), tissues and tissue sections. We can perform transfections and cell culture maintenance. The types of imaging services we provide range from simple 2PPM image acquisition and processing, to more sophisticated experiments involving physiology (timed addition of agonists/inhibitors) or electrophysiology (el. field stimulation, single cell electrophysiology). Our microscopy setup also allows performing 2PPM observations in combination with other imaging modalities (FRET, FRAP).
Experiments can be performed either completely by our microscopy specialists and staff trained in cell culture techniques, or with assistance from visiting scholars. Facilities for accommodating visiting scientists are available.
