Project Details
Projekt
A deconvolution Light-Sheet Microscope for Mesoscopic Tissue Imaging
Subject Area
Immunology
Term
from 2019 to 2024
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 413969432
Light Sheet Fluorescence Microscopy (LSFM) has become a mainstay for biomedical research. While LSFM can reach superresolution and thereby extremely rapid imaging speeds in single cells or small animals, an equally important realm is the imaging of larger animals, tissue sections or even entire organs with cellular resolution, thereby allowing to quickly reconstruct whole organs in 3-D. This is called mesoscopic imaging. It typically requires the chemical treatment of tissue sections or organs to make them optically transparent, a process termed clearing. Within its microscopy core facility IMCES (https://imces.uk-essen.de) the University Hospital Essen is performing mesoscopic LSFM since many years and has developed advanced imaging in many different organs of experimental animals and also human tissue. Thereby, highly original insights have been gained in diverse areas such as physiological organ function, immune rejection, autoimmunity, stroke, myocardial infarction and cancer development. Also new non-toxic laboratory protocols for organ clearing have been developed which have led to a strong increase in the use of the available instrumentation. We are now reaching the limits of the available technology that inhibit us from achieving a deeper scientific understanding, even though it would principally be possible with our approaches. These limits are centered around suboptimal image quality of our LSFM setup that precludes advanced image analysis and hence biological insight. With a new type of deconvolution LSFM that is applied for, here, it would be possible for the first time to obtain an image quality for mesoscopic LSFM data that allows to apply sophisticated methods of image reconstruction and true deconvolution. This is achieved by fundamental improvements in the optical instrument to obtain precise information on light sheet position and quality and hence the point spread function within the entire illumination zone. In addition, innovative and proprietary image reconstruction algorithms are used to de-noise and deconvolve images as well as to achieve the representation of an extremely large spread of pixel intensities from faint cellular processes to very bright cell bodies in one image that fits human vision. The image quality resulting from this operation is far superior over previous technology and will enable totally new insights into the areas of physiological and pathological organ function. It might also have a profound impact on clinical diagnostics, at least of small human organs such as lymph nodes.
DFG Programme
Major Instrumentation Initiatives
Major Instrumentation
High Resolution Microscope
Instrumentation Group
5090 Spezialmikroskope
Applicant Institution
Universität Duisburg-Essen
Participating Person
Dr. Anthony Squire
