Optical trapping with a double laser beam was carried out using a commercial optical tweezer instrument (NanoTracker 2, JPK Instruments, Berlin, Germany), which is constructed on an inverted microscope (IX71, Olympus, Tokyo, Japan) equipped with a CCD camera (DFK 31AF03, The Imaging Source, Taipei, Taiwan). A 1064 nm CW laser beam was split into two beams by a polarization beam splitter, and both beams were focused into the sample through a water-immersed objective lens (60×, N.A. = 1.2) for independent trapping. One of these focal points can be moved by using a piezo-mirror. The laser power at each focal point was set between 40 and 130 mW. All experiments were carried out at room temperature, i.e., 25 °C.
Nanotracker 2
Manufactured by Bruker
Sourced in Germany
The NanoTracker 2 is a high-resolution optical microscope system designed for nanoscale imaging and manipulation. It provides precise control and measurement of single molecules and nanoscale objects.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nanotracker 2
1
Dual Laser Optical Trapping System
2
Probing Dynamics of Cellular Nanotubes
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The details of the experimental setup and necessary procedure were described in our previous report (17 (link)). HeLa cells were transfected to express Lifeact-GFP or Lifeact-mCherry and seeded in PDMS. After more than 24 hours, TAT-coated beads diluted to 4 to 5 fM in cell culture medium were introduced. JPK NanoTracker Desktop Software was used to manage the optical tweezers (NanoTracker 2, JPK Instruments AG) to trap a TAT bead and bind it to a DFB/TNT. At the same time, homebuilt LSCM was used to visualize the fluorescence of FBs to distinguish DFBs from TNTs. The trap stiffness of TAT-coated beads was calibrated to 0.01 pN/nm under our experimental conditions. The trapped and calibrated microspheres were attached to the center of the target a DFB/TNT by moving the stage upward to the trapping focus. After the binding was confirmed by the reduced fluctuation of the axial force signal, the trap was moved orthogonally to the lengthwise axis to deform the linked DFB/TNT (maximum displacement = 3 μm and moving speed = 0.3 μm/s). Measured force signals were analyzed using MATLAB scripts.
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