Ultramicroscope Applications - LaVision BioTec GmbH

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UltraMicroscope II - Applications

Researchers who need artifact free data from overview to a specific region of interest with cellular resolution implement this technology into their projects.
Neuroscientists focusing on the regeneration potential of neurons and the axonal path findinguse this system as do oncologists verifying the efficiency of neovascularization inhibitors. In the field of immunology lymph nodes and the developmental steps of entire lymphatic system are analyzed. The different developmental stages of animal models can be imaged for phenotyping or characterization of pathologies. The image acquisition in vivo is also possible as is the imaging of samples prepared by any clearing procedure. Tissue with endogenous fluorescent proteins like GFP or stained with labelled antibodies can be analyzed fast and easily with this setup. Clearing procedures like 3DISCOor CUBIChave been developed with the UltraMicroscope.


LaVision BioTec reports on the use of light sheet microscopy in the research as part of the Miami Project to Cure Paralysis
LaVison BioTec, developers of advanced microscopy solutions for the life sciences, report on users of their Ultramicroscope Light Sheet Fluorescence Microscope system to aid the research of the Miami Project to Cure Paralysis under the supervision of Professor Vance Lemmon, the Walter G. Ross Distinguished Chair in Developmental Neuroscience & Professor of Neurological Surgery at the University of Miami.

In 2003, Professor Vance Lemmon accepted a position at The Miami Project to Cure Paralysis at the University of Miami. This centre has taken the philosophy that by promoting interactions between basic and clinical scientists, it will be possible to speed the finding of a cure for a devastating clinical problem. This research has focused on answering questions that help define human spinal cord injury and reveal strategies for the repair of damaged spinal tissue.
Professor Lemmon’s lab studies nerve regeneration in the brain and spinal cord. Describing his work, he says “We hope to make nerve cells regenerate much better than they normally do. My colleagues study scar formation in the cord, neural cell translation and blood vessel formation after injury while another colleague studies optic nerve regeneration.”
The biggest challenge to Professor Lemmon and his team is the ability to test literally thousands of samples each week in vitro and rapidly translating interesting “hits” into in vivo tests of efficacy. Imaging and analysis many in vivo samples is vital to this work and this led to the introduction of Light Sheet Fluorescence Microscopy (LSFM) to the team. As he says, “We always want to go faster! We needed to dramatically speed up the pace at which we can image large 3D volumes of the brain and spinal cord. Traditional methods are a bottle neck that limit discovery. While epifluorescence and confocal microscopies were useful, using LSFM enabled us to image way faster and much larger volumes can now be visualised.”








Dr Cynthia Soderblom with the LaVision Ultramicroscope light sheet microscope system in the laboratory of Dr Vance Lemmon at the University of Miami.

For a high resolution copy of the image, either right click to download, or contact Jezz Leckenby at Talking Science.
The work is generating several high profile papers in the field of neuroscience. Professor Lemmon outilnes two. “In our group, we have demonstrated the applicability of LSFM for comprehensive assessment of optic nerve regeneration, providing in-depth analysis of the axonal trajectory and pathfinding. Our study indicates significant axon misguidance in the optic nerve and brain, and underscores the need for investigation of axon guidance mechanisms during optic nerve regeneration in adults. Other colleagues have applied genetic lineage tracing, light sheet fluorescent microscopy and antigenic profiling to identify collagen1α1 cells as perivascular fibroblasts that are distinct from pericytes. The results identify collagen1α1 cells as a novel source of the fibrotic scar after spinal cord injury and shift the focus from the meninges to the vasculature during scar formation.”


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Report on the Neuro Research of Dr Ali Ertürk from the Institute for Stroke and Dementia Research at LMU Munich on the Human Brain After Trauma
Dr Ali Ertürk has been using light sheet and 2-photon microscopy in  his research for a number of years during work in both the USA (with  Genentech) and currently in Germany in the Klinikum der Universität  München (KUM), of the Ludwig Maximilians University (LMU) in Munich.
He is now Group Leader of the Acute Brain Injury Research Group where  his main interest is in understanding key mechanisms leading to  neurodegeneration after traumatic brain injury. At present, virtually  nothing is known about how the initial trauma alters the brain structure  over months/years and ultimately its function.
Dr Ertürk describes the challenges he faces. “One of the main  struggles in neuroscience in general is the difficulty to accurately  analyze long connections in the brain using tissue sections which  deliver only limited spatial information. We use a novel approach aiming  at mapping the acute and chronic changes in the entire brain caused by  small, well-defined brain lesions. To map the pathological brain, we  utilize cutting-edge imaging techniques including high-resolution 3D  imaging of the entire brain - that we recently developed - and in vivo  2-photon imaging. Subsequently, we screen for novel molecular players  that are altered in chronically affected brain regions to halt secondary  neurological problems.”
Choosing the instrumentation from LaVision BioTec for his laboratory  in Munich started with his experience gained using their first  generation UltraMicroscope while in Genentech's Department of  Neuroscience. He was a member of the development team which discovered a  highly reproducible and versatile clearing procedure called 3D imaging  of solvent-cleared organs, or 3DISCO, which is applicable to diverse  tissues including brain, spinal cord, immune organs and tumors.
This has continued in Munich where he images entire transparent  rodent brains. As Dr Ertürk says, the UltraMicroscope is the only  commercial solution for this type of imaging where he looks at  centimeter lengths of tissue. He also makes use of a two-photon  microscope for higher resolution imaging of transparent organs albeit  with a smaller field of view.


 
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