Aligning the μs-ALEX Setup.
- Author(s): Kapanidis, Achillefs;
- Majumdar, Devdoot;
- Heilemann, Mike;
- Nir, Eyal;
- Weiss, Shimon
- et al.
Published Web Locationhttps://doi.org/10.1101/pdb.prot086421
To achieve single-molecule sensitivity and thus have the ability to detect single diffusing fluorophores, careful alignment of the microsecond-alternating laser excitation (μs-Alex) setup is crucial. The following protocol describes routine alignment for 2c-ALEX (532 nm/635 nm) with spectral windows G(550-620)R(650-750).