Multiphoton laser scanning microscope with FLIM module

微宇宙成像中心是耶拿弗里德里希·席勒大学的一个新的成像设施，其目的是支持微宇宙卓越平衡集群的成像需求。为了补充其产品组合，需要一种高度灵活的显微镜，用于通过或在更大的细胞或组织样品层内使用功能荧光读数来研究分子和细胞结构。已经确定了具有代表性的用户，他们的研究将大大受益于该仪器。他们所描述的项目侧重于研究微生物群落对肠道或肺等有机体功能的作用。器官芯片模型正在建立，但由于需要很高的成像深度，直接在芯片中成像到目前为止还不可能。在样品深处使用光学显微镜具有重要的局限性。由于散射和像差导致信号质量严重恶化，导致亮度、对比度和分辨率下降。然而，由于使用红外光，采用双光子或多光子显微镜可以减少散射。此外，微生物组的变异可能导致疾病状态（如炎症）的变化，从而不仅导致细胞重组和分子重组，还导致细胞和分子相互作用、性质和信号事件的变化。诸如荧光寿命或荧光相关光谱（FCS）或光谱成像等替代读数已经突出了对这些特性的微小变化的灵敏度提高。此外，这种功能读数也将用于研究在实际条件下用于显微镜实验的新型定制染料的可用性。另一个项目侧重于真菌和细菌的交流。这涉及多个子项目，如研究金属耐受性、生物矿物形成和细菌（如链霉菌）的生物膜形成。待测样品，如真菌菌丝体或植物根，需要深层组织光学显微镜和基于硬件的技术来去除自身荧光背景。观察膜的性质，如异质脂质堆积及其对蛋白质功能的影响，需要功能性荧光读数。设备的使用、协调、服务和维护任务由Microverse成像中心组织。

The Microverse Imaging Center is a new imaging facility at the Friedrich Schiller University Jena whose aim is to support the imaging needs of the Cluster of Excellence Balance of the Microverse. In order to complement its portfolio, a highly flexible microscope for the investigation of molecular and cellular structures through or within a larger layer of cellular or tissue samples using functional fluorescence readouts, is needed. Representative users have been identified whose research will greatly benefit from the instrumentation. Their described projects focus for instance on studying the action of microbial communities on the function of organisms such as of the gut or the lung. An organ-on-chip model is being established but imaging directly in the chip was so far not possible due to the high imaging depths required. Employing optical microscopy deep inside samples bears important limitations. Signal quality is severely deteriorated due to scattering and aberrations, leading to loss in brightness, contrast and resolution. Scattering is however reduced employing two- or multi-photon microscopy due to the use of infrared light.In addition, variances in the microbiome may entail changes in disease states such as inflammation, and thus result not only in cellular restructuring and molecular reorganizations but also entail variations in cellular and molecular interactions, properties, and signalling events. Alternative readouts such as fluorescence lifetime or fluorescence correlation spectroscopy (FCS) or spectral imaging have highlighted an increased sensitivity already to minute changes of these properties. Furthermore, such functional readouts will also be employed for studying the usability of novel tailor-made dyes for microscopy experiments under real conditions. Another project focuses on the communication of fungi and bacteria. This involves multiple sub-projects such as the study of metal tolerance, biomineral formation and biofilm formation of bacteria such as streptomycetes. The samples to be tested, like fungal mycelium or plant roots, require deep-tissue optical microscopy and hardware-based techniques for the removal of autofluorescence background. Observation of membrane properties such as heterogeneous lipid packing and its influence on protein functionality requires functional fluorescence readouts.Access to the equipment, coordination, service and maintenance tasks are organized by the Microverse Imaging Center.