Single-Molecule Imaging Setup for TIRF Microscopy
Corresponding Organization : Columbia University
Other organizations : The University of Texas Health Science Center at San Antonio
Variable analysis
- 488-nm laser (Coherent Sapphire, 200 mW)
- 561-nm laser (Coherent Sapphire, 200 mW)
- Fluorescence signals captured by two Andor iXon EMCCD cameras
- Prism-type total internal reflection fluorescence (TIRF) microscope (Nikon)
- Lipid bilayer composition (91.5% DOPC, 0.5% biotinylated-PE, 8% mPEG 2000-DOPE)
- SsDNA substrate (generated using rolling circle replication with a biotinylated primer, a circular M13 ssDNA template, and Phi29 DNA polymerase)
- Biotin-streptavidin linkage to attach ssDNA to the lipid bilayer
- Microfluidic system and syringe pump for sample delivery
- Custom-built shuttering system to avoid signal bleed-through during two-color image acquisition
- Not explicitly mentioned
- Not explicitly mentioned
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