ZEISS LSM 510 META CONFOCAL MICROSOPE: CELL & MOLECULAR BIOLOGY

这个子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。描述(由申请人提供)：本申请申请资金购买蔡司LSM 510Meta共焦显微镜系统。这一新仪器将极大地扩展普林斯顿大学目前的荧光显微镜能力。蔡司LSM 510 Meta系统将允许研究人员对同时标记了多个荧光探针的细胞进行成像，包括荧光染料和发射光谱基本上重叠的蛋白质。普林斯顿大学现有的显微镜使用滤光片和二向色镜来选择特定的发射波长，因此仅限于清晰地区分同一细胞中的两种不同的荧光蛋白质(例如YFP和CFP，或GFP和RFP)。此外，当共定位荧光蛋白成像在移动的粒子上时，会出现空间偏移，因为不同的波长在时间上分开的扫描/曝光中被捕获。这两个问题都导致研究进展的范围和速度受到限制。蔡司LSM 510Meta探测器系统解决了这些问题，方法是使用衍射光栅将发射的光扩散到光探测器阵列上，从而有效地确定样品中每个像素的发射光谱。使用先前测量的参考光谱，计算图像中每个点的每个荧光团发射的光的贡献，从而允许单独成像每个荧光团的空间分布。普林斯顿大学目前正在研究的生物系统的初步结果以及其他地方发表的结果表明，蔡司LSM 510Meta探测器将允许研究人员同时对多达8种不同的荧光蛋白质和染料进行成像。这一新能力将使普林斯顿大学的研究人员能够识别活细胞中的多个细胞室，并跟踪快速移动的细胞内颗粒的运输。目前在普林斯顿进行的研究将极大地受益于Meta系统，包括研究果蝇卵母细胞和胚胎中的mRNA运输和细胞运动，疱疹病毒在神经细胞中的组装和运输，乳腺组织中正常和癌变导管的形态发生，线虫的形态发生，以及酵母中接合的细胞生物学。对这些生物系统的研究将提供与重大人类健康问题有关的宝贵信息，如出生缺陷、病毒感染和癌症。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): This application requests funds to purchase a Zeiss LSM 510 Meta confocal microscope system. This new instrument will greatly extend the current capabilities for fluorescent microscopy at Princeton University. The Zeiss LSM 510 Meta system will allow researchers to image cells simultaneously labeled with multiple fluorescent probes, including fluorescent dyes and proteins whose emission spectra substantially overlap. Existing microscopes at Princeton University use optical filters and dichroic mirrors to select specific emitted wavelengths and therefore are limited to cleanly distinguishing only 2 different fluorescent proteins in the same cell (e.g. YFP and CFP, or GFP and RFP). Furthermore, when colocalizing fluorescent proteins are imaged on moving particles, spatial offsets occur because the different wavelengths are captured in temporally separate scans/exposures. Both problems lead to limitations in the scope and rate of research progress. The Zeiss LSM 510 Meta detector system solves these problems by using a diffraction grating to spread the emitted light over an array of light detectors, effectively determining the emission spectrum for every pixel in the specimen. Using previously measured reference spectra, the contribution of emitted light from each fluorophore is computed for every point in the image, allowing the spatial distribution of each fluorophore to be imaged separately. Preliminary results with biological systems currently under study at Princeton University, and published results from elsewhere, demonstrate that the Zeiss LSM 510 Meta detector will allow researchers to simultaneously image as many as eight different fluorescent proteins and dyes. This new capability will allow Princeton researchers to identify multiple cell compartments in living cells and follow the transport of rapidly moving intracellular particles. Research currently undertaken at Princeton which will be benefited greatly by a Meta system include studies of mRNA transport and cell movement in Drosophila oocytes and embryos, herpes virus assembly and transport in neuronal cells, morphogenesis of normal and cancerous ducts in breast tissue, C. elegans morphogenesis, and the cell biology of conjugation in yeast. Study of each of these biological systems will provide valuable information pertaining to significant human health problems such as birth defects, viral infection and cancer.