Potential of correlative luminescence and electron microscopy for detection and location of individual molecules in cells

0438715 - ÚIACH 2015 DE eng A - Abstrakt
Klepárník, Karel
Potential of correlative luminescence and electron microscopy for detection and location of individual molecules in cells.
20th International workshop on "Single Molecule Spectroscopy and Ultrasensitive Analysis in the Life Sciences". Program and Abstract Book.. PiCoQuant GmbH, 2014. s. 128-128.
[International workshop on "Single Molecule Spectroscopy and Ultrasensitive Analysis in the Life Sciences" /20./. 02.09.2015-05.09.2015, Berlin-Adlershof]
Grant CEP: GA ČR(CZ) GA14-28254S
Institucionální podpora: RVO:68081715
Klíčová slova: electron microscopy * individual molecules * cell
Kód oboru RIV: CB - Analytická chemie, separace

Rapidly growing field of correlative light and electron microscopy (CLEM) overcomes some drawbacks of EM together with a time consuming process of the localization of studied structures. This technique bridges the functionality of LM with the high resolution of EM. The dynamic data from an optical source are correlated with ultrastructural analysis done by EM by rendering an overlay image of the same area of the specimen. In the last few years, CLEM has become a unique tool for the investigation of relations between a structure and its biological function. The possibility to reliably map locations of receptors (or any protein) onto a cellular surface with a nanometer accuracy has initiated a tremendous interest in this technique. Usually, a specific antibody is conjugated with a high electron density luminophore nanoparticle to form a probe for tracing cellular objects. Under LM, the luminescence image shows the regions of interest and/or functional changes. Subsequently, an electron beam reveals examined structures highlighted by the nanoparticle dots with the nanometer resolution. A unique approach takes advantage of cathodoluminescence of quantum dots (QD). Electron beam excites QD similarly as a laser and the luminescence emission is detected using a sensitive light detector in the same device. Our investigation is focused on two transmembrane proteins, Fas receptor and Fas ligand. These membrane receptors switch the external apoptotic pathways typical for surveillance strategies of the immune system.
Trvalý link: http://hdl.handle.net/11104/0242154