Newsletter Spring 2022

The 200kV microscope JEOL JEM-2100F operates now with K2 Summit™ Direct Detection Camera (Gatan, Inc., USA). Direct Detection cameras detect the incoming electrons directly on the sensor, which removes limitations caused by noise induced by the electron scattering associated with the conversion of primary electrons to photons and the energy deposition processes, which are processes needed by standard/conventional (CCD) TEM digital cameras. The K2 Summit 4k x 4k direct detection camera is also unique in its ability to count individual electrons (instead of integrating the analogue signal as in standard digital cameras). Another benefit of this detector is the super-resolution mode which can pinpoint electron landing coordinates with sub-pixel accuracy and extends the effective resolution to 57 megapixels, allowing a larger specimen area to be imaged at the same resolution while using lower magnification. Also, the sensor read-out speed of 400 full frames per second adds to the great benefits of this detector which is now mainly used for visualization of cryo-samples.

The routine 120kV microscope JEOL JEM-1400 Flash, installed in our laboratory in June 2021, is equipped with a Xarosa bottom-mounted CMOS camera (EMSIS GmbH, Germany). The latest CMOS technology used in this camera brings together high speed (30 frames per second in its full 20-megapixel resolution), high sensitivity and high dynamic range imaging (HDRI). The Xarosa CMOS camera is installed on our Jeol JEM-1400 Flash microscope with RADIUS software which provides several superior features including e.g. drift correction, “click-to-center”, smart averaging, easy image stitching, full interactive measurements and the integrated image database. Although it offers advanced functions, the RADIUS software managed to make the image acquisition super user-friendly. The JEM-1400 microscope equipped by this camera is now used for high resolution imaging of cell ultrastructure at room temperature but is ready to be used for electron tomography or for work in cryo-conditions.

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The scanner enables acquisition of highly resolved anatomical images (proton density, T1- and T2- weighted images) in vivo virtually of any selected part of rodents’ body, identification of selected metabolites in the tissue by MR spectroscopy, and diffusion weighted images.

Due to the sequence triggering by (breath and/or heart action) the system enables scanning of the abdominal cavity and even the heart in time-resolved image series.

The scanner has a broadband transmit/receive path; therefore, it enables detection also of other nuclei, 19F (using a rat whole body 1H/19F radiofrequency coil) and 31P (1H/31P surface coil).

Output data are both in an intrinsic data format and in DICOM, therefore, they can be easily uploaded and processed by a standard image processing software.

Main focus of this study was to capture and decipher cellular diversity of the embryonic and adult choroid plexus, which play key role in the production of cerebrospinal fluid throughout life. Using combination of various techniques such as single cell profiling and lineage tracing, this study provides a roadmap for better understanding of age-dependent changes in the tissue physiology and generates novel insights of its developmental trajectory.

In our lab, we focus on the dissection of the role of Wnt signalling pathway in various aspects of vertebrate embryonic development, adult homeostasis as well as in multiple pathological conditions. Recently, we identified novel function of the Wnt signalling in the development of choroid plexus. In this study, we were able to leverage our knowledge of the specific role of the Wnt pathway in the choroid plexus embryogenesis to successfully label and trace fate of cells with active Wnt signaling and reveal their contribution to the developing tissue in vivo. Furthermore, using state-of-the-are in situ hybridization approach, we performed high-resolution transcriptomic profiling of these newly identified cells, thus further clarifying the different aspects of the Wnt pathway activation in the choroid plexus development. In summary, our group helped to confirm the observation from single cell analysis of embryonic choroid plexus and establish novel role of Wnt signaling pathway in the choroid plexus morphogenesis.