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Integrating sphere

Manufactured by PerkinElmer

An integrating sphere is a device used for measuring the total radiant flux or luminous flux of a light source. It consists of a hollow spherical cavity with a highly reflective inner surface, which allows for the uniform distribution of light within the sphere. The core function of an integrating sphere is to provide a uniform and diffuse illumination field, enabling accurate measurements of the total output of a light source.

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5 protocols using integrating sphere

1

Optical Characterization of Mixed Nanoparticles

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The formed particles were observed by field-emission scanning electron microscopy (FESEM, FEI Helios NanoLab G3 UC, ThermoFisher Scientific). An accelerating voltage of 5 kV was used. The EDX spectra recording was also realized by FESEM with a JSM-6700F from JEOL Ltd using a silicon drift detector (SDD) XFlash 5130 from Bruker Nano. In the JSM-6700F, the energy of the exciting electrons was set to 10 keV and the accumulation live time was 200 s in all cases. To prevent charging effects in the SEM, a 5 nm thick carbon layer is deposited on the sample before analysis.
For optical characterization of the mixed particles after laser treatment as well as after the etching procedure, the transmission spectra were determined using a spectrometer (H2000, Ocean Optics) setup with a coupled fiber (wavelength range of 200–1100 nm) under transmitted illumination with a halogen-lamp (400–900 nm emission and illumination diameter 2 mm) and with a UV-vis spectrometer with an integrating sphere of PerkinElmer in the same spectral range (illumination area 1 × 2 mm2). All raw spectra are corrected by the background and light source signal.
The resonance spectra of single pure Au and Ag NPs were calculated depending on different diameters in the wavelength range of 350 to 900 nm using the open source software Mie-Plot (https://www.philiplaven.com/mieplot.htm).
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2

Spectral Light Acclimation Protocol

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Before transfer to light with an incident intensity of 61 µE m−2 s−1, dark-arrested cultures in small flasks were covered with colour filters (Lee Filters, UK): Blood Red (#789), Marius Red (#787), Primary Green (#139), Moonlight Blue (#183) and Special Medium Blue (#363). For the flow cytometry and qPCR experiments, a reduced set of filters was used, consisting of Blood Red, Primary Green and Special Medium Blue (Table 1). The transmittance spectrum of each filter (Supplementary Fig. S2) was measured using a Lambda-650S spectrophotometer equipped with an integrating sphere (PerkinElmer). To reduce the potential confounding effect of light intensity on the spectral composition experiments, an approximate correction was made to compensate for the transmittance in the spectral range of each colour filter. The objective was to provide approximately the same incident light intensity, in the wavelength range of the colour filter, for the colour filter treatment versus the control receiving the full spectrum (Supplementary Fig. S3). The approximate correction was made by changing the illumination to compensate for the transmittance at the wavelength of maximum transmittance of each colour filter. Dark and light controls were included by covering flasks with aluminum foil or leaving them uncovered. All experiments were carried out in triplicate.
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3

Spectral Reflectance Analysis using UV/Vis/NIR Spectrophotometer

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Spectrophotometric analysis was performed using a PerkinElmer UV/Vis/NIR Spectrophotometer LAMBDA™ 1050+ [43 ] fitted with a 150 mm InGaAs Integrating Sphere. To calibrate the device, a Spectralon™ certified reflectance standard was utilized, furnished by Labsphere [44 ]. In order to determine the mean spectral reflectance of each sample, three measurements were assessed. The measurements were conducted within a wavelength range of 300–2500 nm, employing a data interval of 5 nm. Subsequently, the data were gathered using the UV WinLab Software, and the ASTM E 903–20 method was applied for executing the reflectance measurements, in tandem with the use of ASTM G 173–03 to acquire terrestrial solar irradiance values. This protocol enabled the calculation of the SR for each individual sample. In Fig. 2, an overview of the PerkinElmer LAMBDA™ 1050+ Spectrophotometer is shown with details of the Integrating Sphere and the sample compartments, and the software used for data elaboration.

An outside view of the PerkinElmer UV/Vis/NIR Spectrophotometer LAMBDA™ 1050+, with details of the Integrating Sphere compartment, the sample holder and the software interface.

Fig. 2
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4

Comprehensive Thermal and Optical Characterization of Materials

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Thermogravimetric analysis (TGA) was performed using a Shimadzu TGA-50 system, in the temperature range from 25 to 800°C at a heating rate of 10°C min–1, under a static atmosphere of air. Polarized optical microscopy (POM) images were obtained with an Olympus BX41 microscope, using crossed polarizers. Scanning Electronic Microscopy (SEM) images were registered with a Hitachi SU-70 electron microscope. The samples were attached to aluminum stubs using double-sided carbon adhesive tape or carbon glue. Pictures were obtained from the surface and cross-section areas. All the samples were sputter-coated with carbon through an EMITECH K950X Turbo Evaporator, at a single pulse, on an outgassing time of 30 s and an evaporating time of 2 s. Transmission electron microscopy (TEM) images were obtained by a Philips CM200 microscope with an accelerating potential of 100 keV. TEM samples were prepared using a method described by Giasson et al. (1988) (link).
UV-Vis reflectance spectroscopy was carried out using a Perkin-Elmer Lambda 950 UV/Vis/NIR spectrophotometer and a Spectralon integrating sphere (Ø = 150 mm). The freestanding film surface was placed perpendicularly to the incident beam and the spectra were acquired as a function of the incident angle (15° < θ < 60°) with 5° steps, as illustrated in Supplementary Scheme 1.
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5

Nanostructure Morphology Analysis

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The morphology of nanostructures is studied by images acquired by the field emission scanning electron microscope (FESEM) (JEOL). The reflection spectra of samples are measured by the UV-visible spectrometer coupled with an integrating sphere (PerkinElmer). Both the scattering and the specular reflection signal are recorded inside the integrating sphere for non-transparent samples in a single measurement. Therefore, the absorption spectrum A can be calculated as A(λ)= 1 -R(λ), where R represents the reflection spectrum.
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