The reference collagen distribution was detected from unstained histological sections using FTIRI (Hyperion 3000 FTIRI Microscope, Bruker Inc., Billerica, MA, USA; imaging settings: 2 x 2 binning, 5.4 × 5.4 μm2 pixel size, 4 cm-1 spectral resolution, on average 32 acquisitions per pixel). Collagen content and distribution were directly evaluated from the area under the Amide I peak (1590–1720 cm-1) of the spectrum. This approach has been validated for collagen distribution within normal and osteoarthritic AC histological slices7,21,22 .
Hyperion 3000 ftiri microscope
Manufactured by Bruker
Sourced in United States
The Hyperion 3000 FTIRI Microscope is a laboratory instrument designed for Fourier Transform Infrared (FTIR) imaging analysis. The core function of this microscope is to capture infrared spectral data from samples, allowing for the identification and characterization of chemical components within the sample.
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Lab products found in correlation
2 protocols using hyperion 3000 ftiri microscope
1
Collagen Imaging in Articular Cartilage
2
FTIRI Microscopy and Depth-Wise Analysis
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The unstained microscopic sections (thickness = 5 μm) were imaged with Hyperion 3000 FTIRI Microscope (Bruker Inc., Billerica, MA, USA) with the following parameters: binning of 2 x 2, pixel size of 5.4 x 5.4 μm, spectral resolution of 8 cm-1, and 16 acquisitions per pixel. The acquired spectral data were offset-normalized, and the integrated absorbances of the amide I region (1595–1720 cm-1) and the carbohydrate region (985–1120 cm-1) were calculated to indicate the distribution of the collagen and the GAGs, respectively [19 (link),23 (link)]. Depth-wise profiles for FTIRI were generated similarly as in CEμCT analysis with slight modification as after generating the 2D profile from single image, the profile was resampled to 100 datapoints without further processing. The relative differences were calculated from the profiles similarly as for the CEμCT profiles (see. Eq 1 ).
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