Inverted Confocal Laser Scanning Microscope

在这个提案中，我们申请更换一台共聚焦激光扫描显微镜，它现在已经使用了10多年。40多个研究小组使用了该仪器。目前的仪器收录了41篇博士论文、100多篇会议论文和至少35篇同行评议出版物。对出版物的系统监测只在过去四年中进行，这意味着出版物的结果可能要高得多。替代仪器将用于不同的科学背景，如医学、自然科学（动物、植物）和材料科学。除了过多的个人极端多才多艺的背景，一些共同的要求与大多数拟议的研究有关。该仪器将用于二维和三维固定样品（细胞、组织、晶体表面）的多通道（平均3 -4）图像采集。仪器的分辨率必须达到阿贝极限，但不能达到超分辨率。发射技术必须能够微调各种发射光谱的探测范围，即使是在不同条件下改变相同荧光染料的行为。一种传统光电倍增管（PMT）和高灵敏度探测器的混合物。在可见光谱中激发所有主要标准荧光激发标签必须是可能的。大的物理视野，特别是成像组织和表面较少的瓷砖，因此较少的拼接应用。此外，高精度的自动对焦必须到位，以获得最佳的三维（z堆栈）成像。所有这些都需要一个全自动显微镜台。

In this proposal we apply for the replacement of a confocal laser scanning microscope, which now is in use for more than 10 years. More than 40 research groups made use of the instrument. The present instrument served 41 PhD thesises, more than 100 conference contributions and at least 35 peer reviewed publications. Systematic monitoring of publications only was performed within the last four years, which means the outcome of publications is likely to be much higher.The replacement instrument will be used on different scientific backgrounds such as medical, natural science (anima, planta) and material science. Independend from the plethora of individual extremly versatile backgrounds a few common requirements are linked to the majority of all proposed research. The instrument will be used for multi-channel (3 -4 in average) image acquisition of fixed samples (cells, tissue, crystalline surfaces) in 2 and 3 dimensions. The instrument must deliver high-resolution down to Abbe's limitation, but no super resolution. Emission technology must enable to fine tune the detection range of various emission spectra, even from within the same fluorescent dye changing behaviour under different conditions. A mixture of conventional photomultiplying tubes (PMT) and high sensitive detectors. Excitation in the visible spectrum at all major standard fluorescent excitation labels must be possible. Big physical field of view, in particular for imaging tissues and surfaces with less tiles and hence less stitching to apply. Further a high precision auto focus must be in place for best threedimenional (z-stack) imaging. All these requires a fully automated microscope stage.