Spectral images were collected in transflection mode using an infrared microscope (Continuum—Thermo Scientific), coupled with an FTIRMS spectrometer (Nicolet 8700—Thermo scientific), equipped with a liquid N2-cooled mercury cadmium telluride detector. All acquisitions were carried out in air, in the mid-IR spectral range (4000–720 cm−1), with a spectral resolution of 8 cm−1, by means of an 18 × 18 (μm × μm) aperture. Stage control and data collection were performed by means of OMNIC 8.0 software (Thermo-Nicolet, Madison, Wisconsin).
Continuum
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The Continuum is a versatile and high-performance Fourier Transform Infrared (FTIR) Spectrometer. It is designed to provide accurate and reliable infrared spectroscopic analysis for a wide range of applications in research and industrial settings. The Continuum offers a robust and stable optical system, delivering consistent and reproducible results.
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4 protocols using continuum
1
Infrared Microscopy Spectral Imaging
2
FTIR Analysis of Macrophage-Bacteria Interactions
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A FTIR microscope (Thermo-Winslet Continuum, UK) attached to a FTIR spectrophotometer (Thermo Nicolet Nexus, UK) bench was used to analyze samples (silicon wafers). The microscope aperture was set to 200 mm x 200 mm and fitted with a type A MCT detector. The FTIR spectra of macrophages incubated with or without bacteria were made up of 52 scans (Omnic 5.2 software, UK) (n=6).
3
Infrared Spectroscopy of Fibrils
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The IR spectra were recorded on a Fourier-transform IR (FT–IR) spectrometer (Avatar360) that was equipped with an IR microscope (Continuum) (Thermofisher Scientific, MA). The 25 × 25 µm area was measured. Each spectrum was measured by a 1024-accumulation cycle at a spectral resolution of 4 cm−1. A pellet of the fibrils, which was prepared via centrifugation was soaked in the buffer solution before it was sandwiched in two CaF2 windows. The IR spectrum was measured for the pelleted particles while they were still wet. A reference sample containing the solvent was also measured at a spot that was in proximity to the pellet.
4
Infrared Microscopy of Protein Aggregates
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One to 2 μl of each suspension was deposited on a CaF2 window and dried at room pressure and temperature. The protein aggregates were then measured in transmission at 50 μm by 50 μm spatial resolution with an infrared microscope (50 (link)). Depending on its size, it was possible to collect 1 to 20 spectra inside each aggregate. The infrared microscope was a Thermo Scientific Continuum equipped with a mercury cadmium telluride (MCT) detector and a 32× 0.65–numerical aperture Reflachromat objective and matching condenser, coupled to a Thermo Scientific Nicolet 8700 spectrometer with a globar source and KBr beamsplitter. The microscope was operated in dual-path single-aperture mode. Spectra were recorded between 650 and 4000 cm−1 at 2 cm−1 resolution, with Happ-Genzel apodization and Mertz phase correction. Spectra were processed in Omnic 9.2 for automatic atmospheric correction to remove water vapor contribution.
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