Associated Data for Single-Molecule Detection of a Fluorescent Nucleobase Analogue via Multiphoton Excitation Authors: David Nobis, Rachel S. Fisher, Mats Simmermacher, Patrycja A. Hopkins, Yitzhak Tor, Anita C. Jones, Steven W. Magennis . 1. Ensemble Fluorescence Spectroscopy -2P cross section calculations -2P power dependence -quantum yield -lifetimes 2.TD-DFT calculations Geometry optimisations (GeoOpt), the resulting 3D structures (xyz files), and the calculations of the three-photon transition probabilities (3PA; the final probabilites can be found in a table in the end of the files). 3. Microscopy Laser spectra used for figure 1 120419_spec_after_glass_cleanupfilter (grey line in figure 1) 130918_spec_110_150 (red line in figure 1) ############################################### Power series with 1f solution and buffer. Plotted in figure 2 are the average count rates from theses measurements (sample minus buffer). 211118_1f_compressed_concentrated_110_150_series1_#.ASC 211118_buffer_compressed_concentrated_110_150_series1_#.ACS # runs from 1 to 13. The power associated with each number is: 1: 0.6 mW 2: 0.9 mW 3: 1.4 mW 4: 1.8 mW 5: 2.8 mW 6: 3.6 mW 7: 4.3 mW 8: 5.9 mW 9: 6.8 mW 10: 8.6 mW 11: 10.0 mW 12: 13.5 mW 13: 18.2 mW ################################################ FCS data to determine counts per molecule. Count rates used for figure S9: 140119_1f_525nM_15min_1.ASC (displayed in figure 3) 140119_background_1min_1.ASC (background for the file above) 070219_f1_1.25muM_slide_2_2.ASC 070219_f1_2.5muM_slide_2_2.ASC 070219_f1_5muM_slide_1_2.ASC 070219_f1_buffer_slide_1.ASC (background for the three files above) ################################################ Example 10 nM Rh110 FCS curve (used to calculate fraction of bright molecules): 140519_Rh110_2.ASC ################################################ MCS data (all shown in figure 4 and S10): Sample 010519_1f1_bpfilter_0205od_110150pulse_timefilter.txt 300419_1f1_bpfilter_0205od_110150pulse_timefilter.txt Buffer 010519_buffer_bpfilter_0205od_110150pulse_timefilter.txt 300419_buffer_bpfilter_0205od_110150pulse_timefilter.txt ################################################ time resolved measurements with 1f (figure S7) 031218_1f_0.5conc_5mhz_0.3mw_tr1.asc (3P regime) 031218_1f_0.5conc_5mhz_0.15mw_tr1.asc (2P regime) ################################################ Pulse optimization (figure S6): Constant width, varying position: 050219_1f_pulseshaping_2_X_#.ASC Constant position, varying width 060219_1f_pulseshape_spectralW_B_1.ASC X=B background shaped and compressed pulse X=S signal shaped and compressed pulse X=RB background compressed pulse X=RS signal shaped and compressed pulse #: scan step