Chemicals for synthesis were purchased from Sigma-Aldrich or Meryer and used as received without any further purification. Dihydro berberine was prepared according to the reported literature.39 The 1H-NMR spectrum was obtained on a Bruker AV 400 spectrometer. High resolution mass spectra (HRMS) were recorded on a GCT premier CAB048 mass spectrometer operating in MALDI-TOF mode. Ultraviolet-visible (UV-vis) absorption spectra were taken on a PerkinElmer Lambda 25 UV-vis absorption spectrophotometer. Photoluminescence spectra were recorded on a PerkinElmer LS 55 fluorescence spectrometer. The absolute fluorescence quantum yields were measured on a Hamamatsu Absolute Quantum Yield Spectrometer C11347.
Lambda 25 uv vis absorption spectrophotometer
Manufactured by PerkinElmer
The Lambda 25 UV/vis absorption spectrophotometer is a versatile instrument designed for accurate and reliable measurements of light absorption in the ultraviolet and visible light spectrum. It is capable of quantifying the concentration of various analytes in liquid samples by measuring the amount of light absorbed at specific wavelengths.
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4 protocols using lambda 25 uv vis absorption spectrophotometer
1
Synthesis and Spectroscopic Characterization
2
Determining Fluorescence Quantum Yield
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The fluorescence quantum yield of IR-X dyes was determined against the reference fluorophore IR-26 dye with a known quantum yield of 0.5%.The optical absorbance was measured for both IR-X dyes solution and IR-26 dye solution at 808 nm using a PerkinElmer Lambda 25 UV/vis absorption spectrophotometer. Then their fluorescence emission intensity was measured using an Edinburgh F920 fluorescence spectrometer equipped with G8605-23 photodetectors under the same 808 nm excitation. Using the measured optical density (OD) and spectrally integrated fluorescence intensity (F), one can calculate the quantum yield of IR-X dyes according to the following formula, Φx(λ)= Φst (λ)*(Fx/Fst) *[(1-10-ODst(λ))/(1-10-ODx(λ))].
3
Characterization of plCSA-Nanoparticles
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UV-vis absorption spectra were taken on a PerkinElmer Lambda 25 UV-Vis absorption spectrophotometer. Photoluminescence (PL) spectra were recorded with an Edinburgh F900 fluorescent spectrometer. Transmission electron microscope (TEM) images were taken on JEM 100CXII (JEOL). The TEM samples were prepared by placing a drop of plCSA-NPs solution onto a 300-mesh copper grid and then drying the sample at room temperature overnight. The particle sizes and zeta potential of particles were characterized on a Nano-Zetasizer (Nano ZS, Malvern, Malvern Instruments, USA) at 25 °C.
4
Spectral Characterization of Fluorescent Dyes
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UV/vis absorption spectra were measured using a PerkinElmer Lambda 25 UV/vis absorption spectrophotometer. Fluorescence emission spectra of IR-X dyes were measured using an Edinburgh F920 fluorescence spectrometer equipped with R928P and G8605-23 photodetectors. Excitation and emission slits were set to 5 nm. Autofluorescence emission spectra of 5% leaf and mouse tissue homogenates were measured using an Edinburgh F920 fluorescence spectrometer equipped with Hamamatsu R928P and G8605-23 photodetectors. Total fluorescence spectra of simulated leaf/muscle were measured using an Edinburgh F920 fluorescence spectrometer equipped with R928P and G8605-23 photodetectors. In the complex system, IR-808 solution placed in a small 4 × 4 mm cuvette was embedded into 5% leaf/mouse tissue homogenate in a 10 × 10 mm cuvette. The spectral response for NIR-Ia and NIR-Ib window detector were measured, and the quantum efficiency of NIR-Ib detector was similar to NIR-Ia (Figure S1 ).
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