A deconvolution Light-Sheet Microscope for Mesoscopic Tissue Imaging

用于介观组织成像的反卷积光片显微镜

• 批准号: 413969432
• 金额: --
• 依托单位: Institut für Experimentelle Immunologie und Bildgebung
• 依托单位国家: 德国
• 项目类别: Major Instrumentation Initiatives
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/413969432?language=en
• 关键词: deconvolution; Light; Sheet; Microscope; Mesoscopic

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.

光片荧光显微镜（LSFM）已成为生物医学研究的支柱。虽然LSFM可以达到超分辨率，从而在单细胞或小动物中实现极快的成像速度，但同样重要的领域是以细胞分辨率对较大动物，组织切片甚至整个器官进行成像，从而允许快速重建3D整个器官。这就是所谓的介观成像。它通常需要对组织切片或器官进行化学处理，使其光学透明，这一过程称为透明化。在其显微镜核心设施IMCES（https：//www.example.com）内，大学医院埃森多年来一直在进行介观LSFM，并在实验动物和人体组织的许多不同器官中开发了先进的成像技术。imces.uk-essen.de因此，在生理器官功能、免疫排斥、自身免疫、中风、心肌梗死和癌症发展等不同领域获得了高度原创的见解。此外，还开发了新的用于器官清除的无毒实验室方案，这导致可用仪器的使用量大幅增加。我们现在正在达到现有技术的极限，这阻碍了我们实现更深入的科学理解，尽管我们的方法基本上是可能的。这些限制集中在我们的LSFM设置的次优图像质量上，这妨碍了高级图像分析，因此也妨碍了生物学洞察。利用这里应用的新型去卷积LSFM，将有可能首次获得介观LSFM数据的图像质量，其允许应用图像重建和真正去卷积的复杂方法。这是通过光学仪器的根本改进来实现的，以获得关于光片位置和质量的精确信息，从而获得整个照明区域内的点扩散函数。此外，创新和专有的图像重建算法用于对图像进行去噪和去卷积，以及在一个适合人类视觉的图像中实现从微弱细胞过程到非常明亮的细胞体的像素强度的极大扩展的表示。这种操作产生的图像质量远远上级以前的技术，将使人们对生理和病理器官功能领域有全新的认识。它也可能对临床诊断产生深远的影响，至少对淋巴结等人体小器官的临床诊断有深远的影响。