Fig. 3: Thermal evaporation of C545T molecules at concentrations suitable for SMFM.
a In-focus single molecule fluorescence images of vacuum-deposited C545T on glass substrates. Spots overlap for deposition times of 15 min and longer, but diffraction-limited spots from single molecule emission are resolvable when the deposition time is reduced to 5 min. Scale bar, 2 µm. For visualisation purposes only, a uniform background was subtracted to improve contrast. b Number of emissive spots per field of view ( ~ 40 × 40 µm2) as a function of C545T deposition time. The number of resolvable spots saturates at long times, as emission from neighbouring molecules begins to overlap. The red shaded area indicates the target concentration range. The grey dashed line is shown as a guide to the eye. A deposition time of 0 min corresponds to substrates that were placed inside the chamber but remained covered during the whole process using the adjustable shutters.
