The OT-AFM Combi-System pairs the exceptional surface force measurement and imaging capabilities of AFM with the ability of optical tweezers to apply and measure smallest forces in 3D.
The OT-AFM Combi-System pairs the exceptional surface force measurement and imaging capabilities of AFM with the ability of optical tweezers to apply and measure smallest forces in 3D. The combined setup fulfills the highest demands on mechanical stability, flexibility, and modularity. A specially designed OT-AFM ConnectorStage™ is the key to combining any AFM of the NanoWizard® or CellHesion® family with the NanoTracker™ optical tweezers on a research-grade inverted optical microscope.
The unique combination of 3D positioning, detection, and manipulation provided by OT and the high-resolution imaging and surface property characterization of AFM opens up a whole new spectrum of applications, such as cellular response, cell-cell or cell matrix interactions, immune response, infection or bacterial/virus/nanoparticle uptake processes, and more.
With a multitude of available handles, interaction, and detection sites, OT-AFM significantly extends the range of single-molecule applications.
1. DNA hairpin unzipping (AFM) while the optical trap can be used to suppress (high laser power) or to quantify rotation (low laser power).
2. Scanning of a decorated DNA molecule. The molecule with DNA binding proteins (green) is spanned between two optically trapped beads. A functionalized AFM tip (blue) scans along the molecule and whenever interactions between the DNA-attached proteins and the tip occur, these can be detected in the AFM and OT signals.
3.监测DNA-酶(例如聚合酶,解旋酶)动力学。随着Onestrand连接到光学捕获的粒子,可以跟踪逐步运动。闭环力夹具允许在单链上保持恒定力。
Cellular response, cell-cell or cell matrix interactions, immune response, infection or bacterial/virus/nanoparticle uptake processes are just a few of the examples that can be investigated with JPK’s new state of the art OT-AFM platform. JPKs proven AFM and OT core technologies, combined with fluorescence microscopy, have set the ultimate benchmark for live cell applications.
[1] + [2]: Activation of cells with functionalized beads, parallel AFM measurement. Signaling molecules on the surface of a microparticle are brought in contact with the cell at defined positions and time points.
[5] + [6]:一个机械敏感的细胞由光学捕获粒子施加的周期力刺激。细胞骨架的内部重排改变了细胞的机械性能。这些属性很容易通过AFM方法访问,例如力量映射或JPK的定量成像高级(QI™-Adadanced)。
[3] + [4]: AFM can be used in parallel to monitor changes in the cell structure, e.g. by monitoring mechanical properties throughout the process or by molecular recognition force spectroscopy that investigates the distribution and mechanical behavior of membrane proteins.
通过使用官能化颗粒或改良的微生物触发细胞反应是一种常见方法。可以使用基于AFM的方法研究细胞结构,动力学和机械性能的结果变化。但是,将物体传递到细胞上的特定感兴趣区域非常困难。OT提供了在精确的时间和位置操纵样品并触发蜂窝响应的理想工具。这显着提高了这些研究的吞吐量,灵活性和可重复性。在此应用中,树突状细胞(DC)和调节性T细胞之间信号的影响(treg) on the adhesion of conventional T-cells (T转换) to the same DC is quantified by OT-AFM.
[1]对树突状细胞(DC)和常规T细胞的粘附实验(T转换)。t转换is attached to a tipless cantilever, then approached to the surface-bound DC. The cantilever is pulled up and the adhesion forces are measured. A regulatory T-cell (Treg) is attached to and removed from the DC with optical tweezers to test its influence on the binding strength. [2]+[3] Measurement setup. The optical trap (red cross) moves the Tregwhile adhesion measurements are performed with a cantilever-attached T转换。[4]针对三种情况测量的分离工作。Treg附件减少了DC-T转换interactions. After the Tregis removed, the adhesion level is almost restored. Sample courtesy of Yan Shi, University of Calgary/Tsinghua University, Beijing. The original experiment was designed by Yan Shi et al. (publication in print).
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