Confocal Laser Scanning Microscope

我们的团队是MHH的新成员；因此，我们必须为实验室配备必要的设备，以继续进行尖端研究。我们使用正常、应激和病理条件下的模式生物黑腹果蝇来研究人类疾病的细胞和分子机制，如肌肉营养不良和神经病理疾病。我们研究的主要目的是解释细胞过程和信号通路，以确保生物体的维持和动态平衡，从而防止疾病的发展。共聚焦激光扫描显微镜(LSM)对于我们的研究是必不可少的，它用于产生高达80%的数据。因此，由于要求的LSM将经历我们集团几乎所有的日常工作时间，因此我们有必要购买一台专门用于我们的仪器。此外，这台显微镜将有益于该研究所组织和细胞成像研究所的所有其他小组。首先，我们使用这种显微镜的研究将揭示压力如何启动亚细胞相分离，以及miRNAs如何参与人类的一系列病理过程。这些发现将使基于miRNA的治疗年龄和压力相关疾病的开发成为可能。其次，对诱导成体干细胞生态位形成的保守信号级联以及调节组织形态和分化的机制进行全面分析，将有助于更好地理解器官发生的原理，这是干细胞生物学和再生医学的核心。此外，干细胞在体内植入、成熟和维持的干细胞生态位的生物工程可以为成功操纵有机化合物及其治疗用途提供基础。这里要求的LSM旨在提高数据生产的质量和效率。该显微镜必须满足高分辨率和强大的共焦显微镜的要求，并应使申请者小组能够获得更快的多通道3D成像的可能性。因此，分辨率、速度、多功能性、灵活性和易用性是最重要的要求。一般要求：显微镜必须1)既可用于固定的组织，也可用于活体组织；2)直立方向；3)在透镜下方有足够的空间，该空间必须以灵活的方式使用；4)既有电动Z驱动器又有XY工作台；5)能够以高分辨率和高灵敏度检测至少四个荧光团(例如，DAPI、GFP、dsRed和Cy5)，并且几乎没有光谱干扰；6)在XYZ方向上达到120x120x350 nm的分辨率，对于活细胞成像实验，在XY分辨率为140 nm或更高的情况下，达到至少18帧每秒的速度。重要的是，我们正在进行的项目所需的所有信息都已包括在报价中。

Our group is new to the MHH; hence it is essential that we equip our lab with the equipment necessary to continue performing cutting-edge research. We use the model organism Drosophila melanogaster under normal, stressed, and pathological conditions to study the cellular and molecular mechanisms underlying human diseases such as muscular dystrophy and neuropathological disorders. The main aim of our research is to explain the cellular processes and signaling pathways that ensure the maintenance and homeostasis of the organism and thus prevent the development of diseases. A confocal laser scanning microscope (LSM) is essential to our research, used in generating up to 80% of our data. For this reason, and since the requested LSM will experience usage nearly all of the daily working hours by our group, it is necessary that we acquire an instrument dedicated for our use. In addition, this microscope will be beneficial to all other groups of the institute for tissue and cell imaging. First, our investigations using this microscope will uncover how stress initiates subcellular phase separation and how miRNAs are involved in a wide range of pathologies in humans. These findings will enable the development of miRNA-based therapeutics for age- and stress-related diseases. Second, a comprehensive analysis of the conserved signaling cascades that induce the formation of an adult stem cell niche and the mechanisms that regulate tissue morphology and differentiation will lead to a better understanding of the principles of organogenesis, which is central to stem cell biology and regenerative medicine. Furthermore, bioengineering of stem cell niches for in vivo engraftment, maturation, and maintenance of stem cells could provide the basis for successful manipulation of organoids and their therapeutic use. The LSM requested here is intended to increase the quality and efficiency of data production. The microscope must meet the requirements of a high-resolution and powerful confocal microscope and should give the applicant group access to the possibilities of faster multi-channel 3D imaging. Therefore, resolution, speed, versatility, flexibility, and ease of use represent paramount requirements. General requirements: the microscope must 1) be usable for both fixed and living tissue; 2) be in an upright orientation; 3) possess ample space below the lens, and this space must be usable in a flexible manner; 4) have both a motorized Z-drive and XY stage; 5) be able to detect at least four fluorophores (e.g., DAPI, GFP, dsRed, and Cy5) with high resolution and sensitivity and with little spectral cross-talk; and 6) achieve a resolution in XYZ directions of 120x120x350 nm, and for live cell imaging experiments, achieve a speed of at least 18 frames per second with an XY resolution of 140 nm or better. It is important that all the information required for the projects we are working on has been included in the offer.