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Direct IR absorption imaging can be achieved using photothermal methods where the AFM cantilever tip acts as an optical excitation detector, while simultaneously recording the image topography [ 6, 7].
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Fluorescence excitation and detector settings were the same for imaging transgenic and wild type tissue sections.
The optical properties in a 1.2cm radius area around the axis of excitation (~450 detectors) were estimated by TRS.
This way both the "fluorescence" channels and the "transmittance" channels are set to detect transmitted excitation light, and detector-by-detector sensitivity differences can be accounted for (see below) because every detector should have been exposed to an identical signal in terms of intensity, pulse delay, and pulse dispersion.
Following PCR amplification, the reaction results are read by flowing the droplets past a fluorescence excitation source and detector, like a flow cytometer without the cells.
First, all the FRET images were recorded using identical parameters, including excitation laser intensity, detector gains and magnification.
OPA-derivatized amino acids were detected with fluorescence detector excitation wavelength 340 nm and emission wavelength 440 nm.
Fluorescence signals from Hoechst and CTXb-647 were captured simultaneously with the reflection signal (excitation 488 nm, detector channel set to 475 525 nm) from cholesterol crystals.
Radiolabeled oligosaccharides were detected by a Packard 150 TR flow-scintillation analyser and oligosaccharide-AP derivatives were quantitated using a Jasco FP-2020 Plus fluorescence detector (excitation wavelength 310 nm, emission wavelength 380 nm).
A 10 µL aliquot of the FMOC reaction product was injected onto a Waters Symmetry C18 column (4.6×250 mm; at 40°C) with Waters Symmetry C18 cartridge guard column and chromatographed using a Dionex GS 50 gradient pump (flow rate = 1.0 mL min−1) and Dionex RS 2000 fluorescence detector (excitation wavelength, 266 nm; emission wavelength, 305 nm).
The fluorescent detector excitation and emission wavelengths were set at 264 and 310 nm, respectively.
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