Vladimir Ermolayev

Publications

• 2.02

  Impact points

  Detection of Single Quantum Dots in Model Systems with Sheet Illumination Microscopy.

  Mike Friedrich, Revaz Nozadze, Sandra de Keijzer, Ralf Steinmeyer, Vladimir Ermolayev, Gregory S Harms

  Journal of fluorescence. 10/2011;

  Single molecule detection and tracking provides at times the only possible method to observe the interactions of low numbers of biomolecules, inlcuding DNA, receptors and signal mediating proteins in living systems. However, most existing imaging methods do not enable both high sensitivity and non-i... [more] Single molecule detection and tracking provides at times the only possible method to observe the interactions of low numbers of biomolecules, inlcuding DNA, receptors and signal mediating proteins in living systems. However, most existing imaging methods do not enable both high sensitivity and non-invasive imaging of large specimens. In this study we report a new setup for selective plane illumination microscopy (SPIM), which enables fast imaging and single molecule tracking with the resolution of confocal microscopy and the optical penetration beyond 300 μm. We detect and report our instrumental figures of merit, control values of fluorescence properties of single nano crystals in comparison to both standard widefield configurations, and also values of nanocrystals in multicellular "fruiting bodies" of Dictyostelium, an excellent control as a model developmental system. In the Dictyostelium , we also report some of our first tracking of single nanocrystals with SPIM. The new SPIM setup represents a new technique, which enables fast single molecule imaging and tracking in living systems.
• 4.39

  Impact points

  STED-SPIM: Stimulated emission depletion improves sheet illumination microscopy resolution.

  Mike Friedrich, Qiang Gan, Vladimir Ermolayev, Gregory S Harms

  Biophysical journal. 04/2011; 100(8):L43-5.

  We demonstrate the first, to our knowledge, integration of stimulated emission depletion (STED) with selective plane illumination microscopy (SPIM). Using this method, we were able to obtain up to 60% improvements in axial resolution with lateral resolution enhancements in control samples and zebraf... [more] We demonstrate the first, to our knowledge, integration of stimulated emission depletion (STED) with selective plane illumination microscopy (SPIM). Using this method, we were able to obtain up to 60% improvements in axial resolution with lateral resolution enhancements in control samples and zebrafish embryos. The integrated STED-SPIM method combines the advantages of SPIM with the resolution enhancement of STED, and thus provides a method for fast, high-resolution imaging with >100 μm deep penetration into biological tissue.
• 2.55

  Impact points

  Detection of single quantum dots in model organisms with sheet illumination microscopy.

  Mike Friedrich, Revaz Nozadze, Qiang Gan, Monika Zelman-Femiak, Vladimir Ermolayev, Toni U Wagner, Gregory S Harms

  Biochemical and biophysical research communications. 10/2009;

  Single-molecule detection and tracking is important for observing biomolecule interactions in the microenvironment. Here we report selective plane illumination microscopy (SPIM) with single-molecule detection in living organisms, which enables fast imaging and single-molecule tracking and optical pe... [more] Single-molecule detection and tracking is important for observing biomolecule interactions in the microenvironment. Here we report selective plane illumination microscopy (SPIM) with single-molecule detection in living organisms, which enables fast imaging and single-molecule tracking and optical penetration beyond 300mum. We detected single nanocrystals in Drosophila larvae and zebrafish embryo. We also report our first tracking of single quantum dots during zebrafish development, which displays a transition from flow to confined motion prior to the blastula stage. The new SPIM setup represents a new technique, which enables fast single-molecule imaging and tracking in living systems.
• 8.98

  Impact points

  Impaired axonal transport in motor neurons correlates with clinical prion disease.

  Vladimir Ermolayev, Toni Cathomen, Julia Merk, Mike Friedrich, Wolfgang Härtig, Gregory S Harms, Michael A Klein, Eckhard Flechsig

  PLoS pathogens. 09/2009; 5(8):e1000558.

  Prion diseases are fatal neurodegenerative disorders causing motor dysfunctions, dementia and neuropathological changes such as spongiosis, astroglyosis and neuronal loss. The chain of events leading to the clinical disease and the role of distinct brain areas are still poorly understood. The role o... [more] Prion diseases are fatal neurodegenerative disorders causing motor dysfunctions, dementia and neuropathological changes such as spongiosis, astroglyosis and neuronal loss. The chain of events leading to the clinical disease and the role of distinct brain areas are still poorly understood. The role of nervous system integrity and axonal properties in prion pathology are still elusive. There is no evidence of both the functional axonal impairments in vivo and their connection with prion disease. We studied the functional axonal impairments in motor neurons at the onset of clinical prion disease using the combination of tracing as a functional assay for axonal transport with immunohistochemistry experiments. Well-established and novel confocal and ultramicroscopy techniques were used to image and quantify labeled neurons. Despite profound differences in the incubation times, 30% to 45% of neurons in the red nucleus of different mouse lines showed axonal transport impairments at the disease onset bilaterally after intracerebral prion inoculation and unilaterally -- after inoculation into the right sciatic nerve. Up to 94% of motor cortex neurons also demonstrated transport defects upon analysis by alternative imaging methods. Our data connect axonal transport impairments with disease symptoms for different prion strains and inoculation routes and establish further insight on the development of prion pathology in vivo. The alterations in localization of the proteins involved in the retrograde axonal transport allow us to propose a mechanism of transport disruption, which involves Rab7-mediated cargo attachment to the dynein-dynactin pathway. These findings suggest novel targets for therapeutic and diagnostic approaches in the early stages of prion disease.
• 4.39

  Impact points

  Ultramicroscopy reveals axonal transport impairments in cortical motor neurons at prion disease.

  Vladimir Ermolayev, Mike Friedrich, Revaz Nozadze, Toni Cathomen, Michael A Klein, Gregory S Harms, Eckhard Flechsig

  Biophysical journal. 05/2009; 96(8):3390-8.

  The functional imaging of neuronal circuits of the central nervous system is crucial for phenotype screenings or investigations of defects in neurodegenerative disorders. Current techniques yield either low penetration depth, yield poor resolution, or are restricted by the age of the animals. Here, ... [more] The functional imaging of neuronal circuits of the central nervous system is crucial for phenotype screenings or investigations of defects in neurodegenerative disorders. Current techniques yield either low penetration depth, yield poor resolution, or are restricted by the age of the animals. Here, we present a novel ultramicroscopy protocol for fluorescence imaging and three-dimensional reconstruction in the central nervous system of adult mice. In combination with tracing as a functional assay for axonal transport, retrogradely labeled descending motor neurons were visualized with >4 mm penetration depth. The analysis of the motor cortex shortly before the onset of clinical prion disease revealed that >80% neurons have functional impairments in axonal transport. Our study provides evidence that prion disease is associated with severe axonal transport defects in the cortical motor neurons and suggests a novel mechanism for prion-mediated neurodegeneration.
• 4.27

  Impact points

  Comparison of Al-induced gene expression in sensitive and tolerant soybean cultivars.

  Vladimir Ermolayev, Winfriede Weschke, Renate Manteuffel

  Journal of experimental botany. 01/2004; 54(393):2745-56.

  In order to identify genes involved in soybean resistance to aluminium (Al) stress differential gene expression patterns of Al-stressed and non-stressed tolerant and sensitive soybean cultivars were compared. Out of eight described genes, potentially related to mechanisms of aluminium stress, only p... [more] In order to identify genes involved in soybean resistance to aluminium (Al) stress differential gene expression patterns of Al-stressed and non-stressed tolerant and sensitive soybean cultivars were compared. Out of eight described genes, potentially related to mechanisms of aluminium stress, only phosphoenolpyruvate carboxylase (PEPC) revealed enhanced expression in roots of tolerant as compared to sensitive soybean cultivars under stress conditions. Additionally, two novel full-length cDNA sequences, homologous to translationally controlled tumour proteins (TCTP, clone 58, GenBank accession number AF421558) and inosine-5'-monophosphate dehydrogenases (IMPDH, clone 633, GenBank accession number AF421559) with enhanced expression of the corresponding genes only in roots of Al-tolerant soybean cultivar under stress conditions were isolated and characterized. For functional analysis full-length cDNA 633 was transferred in Arabidopsis thaliana. Only 6% of the seedlings from the wild type survived Al stress, whereas 86% of transgenics were vital demonstrating superiority in stress protection. Compared with the wild type, transgenic plants showed diminished Al penetration into the roots after the stress treatment especially in the division and elongation zones of the roots. Formation of numerous lateral roots in transgenic plants with low elicited callose accumulation under stress conditions indicated ability of the IMPDH homologue to mediate aluminium tolerance in transgenic plants. Possible functional activities of Al up-regulated genes in resistance mechanisms are discussed.