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Re-opening experiments was measured at 30°C using a SpectraMax5 plate reader with excitation and emission wavelengths as above and with a 540 nm emission cutoff.
The particular numbers shown in Figure 1 were achieved by fixing the posterior and transition cutoffs to 0.6 and 10 − 5, respectively, and setting the emission cutoff to 10 − 5, 10 − 4, 10 − 3, 10 − 2, 0.05, 0.1, 0.4 and 0.99.
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The settings for fluorescence were excitation 480 nm, emission 510 nm, cutoff filter at 495 nm, PMT automatic.
Fluorescent product, caused by hydrogen peroxide-dependent, Amplex Red oxidation during hDAAO-catalysed substrate turnover, was measured on a FlexStation II (Molecular Devices) in endpoint mode with these settings: excitation 530 nm, emission 590 nm, cutoff 590 nm and photomultiplier tube on Low.
(i) In the first scheme, multielectron/multistep excitation processes should be introduced; however, these multiexcitation processes must be excluded because of the low quantum efficiency of inelastic tunneling [32], which is also supported by the sample bias dependence of the STM-LE results (Figure 6) in which all of the emissions satisfied the cutoff condition (hν ≤ eVs).
When emission was below this cutoff the quantum dot was defined as residing in a "dark" or "off" state.
Fluorescence intensity was recorded at 535 nm emission (with 530 nm cutoff filter) using 420 nm and 500 nm excitation filters.
Substrate hydrolysis rate was followed in a Spectramax Gemini XPS 96-well plate fluorescence reader (Molecular Devices) using 365 nm excitation and 450 nm emission wavelength with a cutoff at 435 nm.
The 488 nm laser line was used with <4% of transmission power, and the eGFP signal was detected with a long-pass emission filter (505 nm cutoff).
An emission short-pass filter (cutoff 690 nm) was placed in the microscope scanhead to avoid background irradiance from the excitation source.
Fluorescence was measured with a Gemini XS spectrofluorometer with 1 min intervals for 1 h at 40°C using ddH2O as blank, with the following parameters: excitation 330 nm, emission 456 nm, and cutoff 455 nm.
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