BioRam 活体单细胞显微激光拉曼光谱仪
拉曼散射技术与荧光光谱与红外光谱等技术相比,不仅具有可在无须荧光标记情况下对活细胞进行基本无扰、原位测定的优点,而且拉曼光谱不受水的干扰。故拉曼光谱技术在研究测定活细胞尤其是连续监测单个活细胞上有无可比拟的突出优点。
BioRam 活体单细胞显微激光拉曼光谱仪可在无任何标记情况下对活细胞进行三维扫描测定其分子分布,更具优越性。BioRam 活体单细胞显微激光拉曼光谱仪为医师，药师和生物学家提供了利用拉曼光谱的便利的工具，BioRam系统中显微镜，拉曼光谱和特殊定制软件的创新组合，不仅可以识别各种基团，还可以对化合物进行高精确度分析，且对活细胞不需要使用生化标记物，荧光标记物或者抗体，对活细胞安全。 系统亮点概述
（1）对于细胞活力和培养性没有限制 相比于利用FACS或MACS原理来进行细胞识别的仪器，BioRam拉曼光谱仪无需使用磁珠，生化标记物或荧光标记物，可在整个过程中保持细胞的活性。 测试后的细胞可以传代培养以用于进一步的实验。在这种过程中，由于细胞的活性保持不变，所以可以对癌细胞的特征进行分析，并可以对它们与各种活性物质相互作用的效果进行分析。BioRam的应用范围涵盖从生物研究到临床实践，可对病人的具体治疗决策进行支持。 
图2 对细胞表面氨基酸的拉曼指纹图谱分析 （2）即使在液体环境中，也可对细胞进行精确分析
并非所有的细胞都具有清晰可辨的表面标志物。但在区分癌细胞与健康细胞、或干细胞等具有分化能力的细胞过程中，Rarnan光谱显著优于其它方法。BioRam技术可以对细胞表面基团标志物进行精确分析，例如对于红细胞，白细胞，急性髓性白血病细胞或乳腺癌细胞等[2,3]。所以该技术可以对未定影的组织或活检材料，如致病血液细胞等进行诊断快速和可靠地识别。同样，干细胞也可以应用这种原理从外周血或脐带血中分离出来，以用在对它们的自然状态的研究或治疗方法中。 图3 对于不同血液细胞的拉曼光谱分析 （3）测量技术简单易行
样品不用特别制备。可以直接使用活体组织或在水溶液中对细胞进行测量。同样，细胞可以放在与拉曼光谱仪兼容的培养皿中或者微流控芯片中分析。直接“点击运行”即可获取高质量光谱数据，所产生的光谱与数据库比较可以分析细胞类型和状态。同时，原始数据和所有显微镜和激光的设置参数也被保存，为测量的后续实验提供依据，以进行进一步的分析。 （4）允许培养组织进行非侵入性分析
该BioRam系统允许培养组织进行非侵入性分析。例如，在自体软骨移植的软骨细胞的生存能力和纯度以及它们的分化状态可用BioRam系统进行Raman光谱的测试[4]。此外利用BioRam系统对人类皮肤成纤维细胞，角质形成细胞，黑素细胞和角质形成细胞在活体条件下进行的分析[5]，在间充质干细胞的培养过程中，基质成纤维细胞的污染也被证实具有高度的特异性 [6]。这开辟了许多可能的应用领域，例如在在细胞培养和组织工程中监测和质量控制。 AdvantagesRapid, exact analysis and new applicationsCellTool’s platforms for the photonic analysis of biological samples have significant advantages over established techniques such as FACS (fluorescence activated cell sorting), MACS (magnetic activated cell sorting) or immunohistochemistry. They offer completely new opportunities to physicians, pharmacists and
biologists in many applications. Find a summary of key advantages below: Unaltered and viable cells, available for downstream applicationsNo labelling with biochemical markers, colours, antibodies or beads No modification of cell surfaces or activity states No physical stress through shear forces or non-physiological conditions No destruction of biological material Perform multiple consecutive analyses on the same cell or tissue! Use cells or tissues subsequently for other applications! Monitor cell or tissue cultures online without loosing cells or tissues!
Precise results, even in liquidsResults are no longer limited to the markers you use Very few cells deliver unequivocal results, even in liquids Identify cell types and activity states, even when no appropriate marker is available! Clearly distinguish different stem cell types and cancerous from healthy cells! Determine the differentiation stage of stem cells! Spot rare cells in human samples with high sensitivity! Investigate cellular interactions and the effects of small molecules in vivo!
Broad range of samples, small volumesAll kinds of microorganisms, cell-lines, primary cells, tissues and tissue sections Volumes in the ?l to a few ml range Analyse even minimal (e.g. clinical) samples reliably!
Comprehensive, easy-to-use systemsTailored platforms for various applications Fully integrated components and automated processes, from mounting to measurement and data analysis Add-on modules available, e.g. optical ‘teasers’ for deeper cell analysis or microfluidics for automation, increased throughput and sorting of cells Intuitive user interface First-class customer support Workshops and application laboratory for sharing knowledge and exploring new ideas Just get going and join our expert community! 3、适用范围
在没有标记物和生理条件下，对各种微小有机体，细胞系，原代细胞和组织进行拉曼光谱分析，确定细胞类型和活动状态； 区分不同的干细胞类型或确定它们的分化阶段； 监测并记录在细胞和组织培养细胞过程中的变化； 可以识别非常少见的细胞，对人体标本具有较高的特异性分析能力； 在无剪切力、无生理上的压力情况下对样品分析，使细胞保持活力利于下游应用。 分析培养的细胞和活性物质之间的相互作用。
图4 BioRam拉曼光谱仪可监测并记录在细胞培养过程中的变化 ApplicationsA paradigm shift in the identification, characterization and sorting of cellsCellTool’s analytical platforms enable physicians, pharmacists and biologists to exploit the potential of Raman spectroscopy for biomedical applications. Just like ‘photonic fingerprinting’ Raman spectroscopy provides specific information about the overall chemical composition of the specimen. It enables reliable identification of different cell types, discrimination of cell phase and even monitoring of stem dell differentiation without any biochemical labeling such as antibody-based markers or fluorescence molecules. Browse our selection of examples ranging from cancer research and stem cell research to drug discovery , quality control and clinical practice. In cooperation with partners, we are continuously exploring further applications and expanding the body of knowledge in the field. We engage in an open dialogue with academia, clinics and industry and expressly welcome your input. 4、参数 BioRam光谱仪——
激光器：785 nm激光器
通光效率大于30%
光谱范围： 100nm-3400nm，光谱分辨率：1cm-1
空间分辨率：横向0.5微米，纵向2微米
低波数：200 cm-1，100 cm-1，50 cm-1，10 cm-1供选择
软件：应用于PC或Mac的创新软件，自动化系统控制（包括电激光控制快门联锁系统及配置的用户界面）
计算机及主机——
CPU：1GHZ以上；内存：500MB 以上；硬盘 80G以上
5、使用方法
（1）开机
1、打开主机电源；
2、计算机电源
3、将使用的激光器电源
（2）自检
1、用鼠标双击图标，进入仪器工作软件环境；
2、系统自检画面出现，选择仪器并确定。系统将检验所有的激光器、电机。
（3）实验
1、实验条件设置
2、观察和采谱：执行测量命令。
（4）关机
1、关闭计算机
2、关闭主机电源；
3、关闭激光器参考文献
(1) Evaluierung der Raman SpeMroskopie f~r die marker- und zerst~rungsfreie Qualit~tskontrolle im Tissue Engineering. Steffen Koch (2010) Fraunhoferverlag ISBN: 978-3-8396-0112 9
{2) Nondestructive Identification of Individual Leukemia Cells by Laser Trapping Raman Spectroscopy. James W. Chan et al. Anal. Chem. (2OO8) 80:2180 2187
{3) Turnour cell identification by means of Rarnan spectroscopy in combination with optical traps and rnicrofiuidic environments. Sebastian Dochow et al. Lab Chip (2011) 11:1484
(4) Raman spectroscopy as a tool for quality and sterility analysis for tissue engineering applications like cartilage transplants. Marieke Pudlas et al. Int J Artif Organs (2010) 33,3:228 237
(5) Rarnan Spectroscopy: A Noninvasive Analysis Tool For The Discrimination of Human Skin Cells. Marieke Pudlas et al. Tissue Engineering (2011) C 17,10:1027 1040
{6) Non-contact discrimination of human bone marrow-derived mesenchymal stem cells and fibroblasts using Raman spectroscopy. Marieke Pudlas et al. Medical Laser Application (2011) 26:119 125 图像参考
Page 2: Osteosarcorna cells Steffen Koch, Fraunhofer IGB Stuttgart, Germany
Page 3: Rarnan spectra Steffen Koch (2010) Fraunhoferverlag ISBN: 978-3-8396-0112-9, Germany
Page 4: Laser guidance for Rarnan spectroscopy Carsten Bolwien, Fraunhofer IPM Freiburg, Germany
Page 6: Spectra of various blood cells Steffen Koch, Fraunhofer IGB Stuttgart, Germany
Page 7: Graft from autologous cartilage biopsy material, before and after transplantation Neike Walles, Fraunhofer IGB Stuttgart, Germany
Page 7: Typical Rarnan spectra of rnesenchyrnal stern cells and osteosarcorna cells with cluster plot after statistical analysis Marieke Pudlas, Fraunhofer IGB Stuttgart, Germany
Page 9: Mesenchyrnal stern cells with laser beam Steffen Koch und Carsten Bolwien, Germany |
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