OT-AFM Combi-System

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.

三维定位的独特组合,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. Monitoring of DNA-enzyme (e.g. polymerase, helicase) dynamics. With onestrand attached to an optically trapped particle, the step-wise motion can be tracked. Closed-loop force clamping allows maintaining a constant force on the single strand.

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]: A mechano-sensitive cell is stimulated by a periodic force, exerted by an optically trapped particle. Internal rearrangements of the cytoskeleton alter the mechanical properties of the cell. These properties are easily accessed with AFM methods like force mapping or JPK’s Quantitative Imaging Advanced (QI™-Advanced).

[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（t_reg) on the adhesion of conventional T-cells (T_转换) to the same DC is quantified by OT-AFM.

[1] Adhesion experiment with dendritic cells (DC) and conventional T-cells (T_转换). The 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 (T_reg) 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 T_regwhile adhesion measurements are performed with a cantilever-attached T_转换. [4] Detachment work measured for the three situations. Treg attachment reduces DC-T_转换interactions. After the T_regis 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).

• Imaging, positioning, and manipulation experiments from single molecules to living cells
• Measure forces in 2D and 3D, from 500fN to 10nN, on the same sample
• Fully flexible and modular design, with the widest range of modes & accessories
• 与光学显微镜技术（例如TIRF或共聚焦）的全面整合
• All NanoTracker™, NanoWizard® /CellHesion® 200 systems can be combined to become a complete OT-AFM-platform
• ConnectorStage™将AFM与Tweezers硬件结合在所有主要倒置的光学显微镜上，来自Zeiss，Nikon，Olympus和Leica

• All NanoTracker™, NanoWizard®/CellHesion® 200 systems can be combined to become a complete OT-AFM-platform
• ConnectorStage™可以将AFM与Tweezers硬件结合在所有主要的倒置光学显微镜上，来自Zeiss，Olympus，Nikon，Leica

• Two separate controllers drive the two systems independently:
  
  
  • NanoWizard® AFM controller with software
  • NanoTracker™ OT controller with software

Optical systems/accessories, electrochemistry solutions, electrical sample characterization, environmental control options, software modules, temperature control, acoustic and vibration isolation solutions and more. Bruker provides you with the right accessories to control your sample conditions and to perform successful experiments.