ZEISS LSM 510 META CONFOCAL MICROSOPE: CANCER

该子项目是利用由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。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（由申请人提供）：本申请要求资金购买蔡司LSM 510 Meta共聚焦显微镜系统。这种新仪器将大大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有多种荧光探针的细胞进行成像，包括荧光染料和发射光谱基本重叠的蛋白质。普林斯顿大学现有的显微镜使用光学滤波器和分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（例如YFP和CFP，或GFP和RFP）。此外，当共定位荧光蛋白在移动粒子上成像时，由于在时间上分离的扫描/曝光中捕获不同的波长，因此发生空间偏移。这两个问题导致研究进展的范围和速度受到限制。Zeiss LSM 510 Meta检测器系统通过使用衍射光栅将发射光传播到光检测器阵列上来解决这些问题，从而有效地确定样本中每个像素的发射光谱。使用先前测量的参考光谱，针对图像中的每个点计算来自每个荧光团的发射光的贡献，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 510 Meta检测器将允许研究人员同时成像多达8种不同的荧光蛋白和染料。这种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区室，并跟踪快速移动的细胞内颗粒的运输。目前在普林斯顿大学进行的研究将大大受益于Meta系统，包括果蝇卵母细胞和胚胎中mRNA运输和细胞运动的研究，神经元细胞中疱疹病毒的组装和运输，乳腺组织中正常和癌性导管的形态发生，C.线虫形态发生和酵母接合的细胞生物学。对这些生物系统中的每一个系统的研究将提供与重大人类健康问题有关的有价值的信息，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.