Figure 6. X-ray microprobe analysis of iron in the T. rex vascular tissues. (a,b,e) Optical micro... more Figure 6. X-ray microprobe analysis of iron in the T. rex vascular tissues. (a,b,e) Optical microscope images of vessel tissues and (c,d,f) corresponding iron µ-XRF distribution maps recorded at 10 keV. Brighter pixels correspond to higher Fe content. All scale bars are 50 µm. Additional elemental maps of regions (a) and (b) can be found in Fig. S5. In (b,d) the vessel structure is not an organic tissue but a mineralised cast rich in Ba and S (see Fig. S5). Such fine-scale variation in preservation underscores the notion that preservation depends on the microenvironment. Numbered white circles indicate locations of Fe µ-XANES analysis. (g) Stacked normalised Fe K-edge extended XANES spectra of spots 0–6. Fits are shown in red dashed lines, with corresponding residuals plotted at the bottom. All spectra match to goethite (α-FeO(OH)) with normalised sum-square values ranging from 0.59 to 1.93·10−4. For comparison, an example set of the iron bearing reference spectra used are displayed...
Figure 2. Microscopy images of T. rex vascular tissue and associated analysis of fibrillar collag... more Figure 2. Microscopy images of T. rex vascular tissue and associated analysis of fibrillar collagen banding. (a) Transmitted VLM of T. rex soft tissue shows an extensive network of hollow, pliable, vascular structure and typical brown hue. (b) SEM image of the surface of a vessel. (c) Magnified image of (b) detailing features consistent with collagen fibre bundles (collagen fibril, "f "; collagen fibre, "CF"). Average fibril width was measured as 110 nm, and average fibre width, 1.0 µm. (d) TEM image of fibrous features observed in a longitudinal vessel cross-section. Intensity profiles of banded texture in (e) boxes 1 and 2 in c and (f) boxes 3, 4, 5 in (d) with example peak-to-peak distances (SEM average, ~74 nm; TEM, ~56 nm) called out in red. See Fig. S6 for precise d-spacing values determined using SAXS. For comparison to a modern blood vessel network in bone, see Fig. 5b of ref.39.
Figure 3. SR-FTIR full spectra of isolated T. rex vascular tissue and chicken type I collagen (no... more Figure 3. SR-FTIR full spectra of isolated T. rex vascular tissue and chicken type I collagen (no treatment). All key bands for the identification of protein (Amide I, Amide II, Amide III) are present in the dinosaur tissue spectrum. The T. rex spectrum also presents a strong non-peptide carbonyl (C=O) band at 1739 cm−1 and a carbohydrate band at ~1010 cm−1.
Author(s): Holman, Hoi-Ying N.; Comolli, Luis R.; Wozei, Eleanor; Hazen, Terry C.; Downing, Kenne... more Author(s): Holman, Hoi-Ying N.; Comolli, Luis R.; Wozei, Eleanor; Hazen, Terry C.; Downing, Kenneth H.
Water is a strong mid-infrared absorber, which has hindered the full exploitation of label-free a... more Water is a strong mid-infrared absorber, which has hindered the full exploitation of label-free and non-invasive infrared (IR) spectromicroscopy techniques for the study of living biological samples. To overcome this barrier, many researchers have built sophisticated fluidic chambers or microfluidic chips wherein the depth of the liquid medium in the sample compartment is limited to 10 μm or less. Here we report an innovative and simple way to fabricate plastic devices with infrared transparent view-ports enabling infrared spectromicroscopy of living biological samples; therefore the device is named "IR-Live". Advantages of this approach include lower production costs, a minimal need to access a micro-fabrication facility, and unlimited mass or waste exchange for the living samples surrounding the view-port area. We demonstrate that the low-cost IR-Live in combination with microfluidic perfusion techniques enables long term (>60 h) cell culture, which broadens the capab...
Figure 6. X-ray microprobe analysis of iron in the T. rex vascular tissues. (a,b,e) Optical micro... more Figure 6. X-ray microprobe analysis of iron in the T. rex vascular tissues. (a,b,e) Optical microscope images of vessel tissues and (c,d,f) corresponding iron µ-XRF distribution maps recorded at 10 keV. Brighter pixels correspond to higher Fe content. All scale bars are 50 µm. Additional elemental maps of regions (a) and (b) can be found in Fig. S5. In (b,d) the vessel structure is not an organic tissue but a mineralised cast rich in Ba and S (see Fig. S5). Such fine-scale variation in preservation underscores the notion that preservation depends on the microenvironment. Numbered white circles indicate locations of Fe µ-XANES analysis. (g) Stacked normalised Fe K-edge extended XANES spectra of spots 0–6. Fits are shown in red dashed lines, with corresponding residuals plotted at the bottom. All spectra match to goethite (α-FeO(OH)) with normalised sum-square values ranging from 0.59 to 1.93·10−4. For comparison, an example set of the iron bearing reference spectra used are displayed...
Figure 2. Microscopy images of T. rex vascular tissue and associated analysis of fibrillar collag... more Figure 2. Microscopy images of T. rex vascular tissue and associated analysis of fibrillar collagen banding. (a) Transmitted VLM of T. rex soft tissue shows an extensive network of hollow, pliable, vascular structure and typical brown hue. (b) SEM image of the surface of a vessel. (c) Magnified image of (b) detailing features consistent with collagen fibre bundles (collagen fibril, "f "; collagen fibre, "CF"). Average fibril width was measured as 110 nm, and average fibre width, 1.0 µm. (d) TEM image of fibrous features observed in a longitudinal vessel cross-section. Intensity profiles of banded texture in (e) boxes 1 and 2 in c and (f) boxes 3, 4, 5 in (d) with example peak-to-peak distances (SEM average, ~74 nm; TEM, ~56 nm) called out in red. See Fig. S6 for precise d-spacing values determined using SAXS. For comparison to a modern blood vessel network in bone, see Fig. 5b of ref.39.
Figure 3. SR-FTIR full spectra of isolated T. rex vascular tissue and chicken type I collagen (no... more Figure 3. SR-FTIR full spectra of isolated T. rex vascular tissue and chicken type I collagen (no treatment). All key bands for the identification of protein (Amide I, Amide II, Amide III) are present in the dinosaur tissue spectrum. The T. rex spectrum also presents a strong non-peptide carbonyl (C=O) band at 1739 cm−1 and a carbohydrate band at ~1010 cm−1.
Author(s): Holman, Hoi-Ying N.; Comolli, Luis R.; Wozei, Eleanor; Hazen, Terry C.; Downing, Kenne... more Author(s): Holman, Hoi-Ying N.; Comolli, Luis R.; Wozei, Eleanor; Hazen, Terry C.; Downing, Kenneth H.
Water is a strong mid-infrared absorber, which has hindered the full exploitation of label-free a... more Water is a strong mid-infrared absorber, which has hindered the full exploitation of label-free and non-invasive infrared (IR) spectromicroscopy techniques for the study of living biological samples. To overcome this barrier, many researchers have built sophisticated fluidic chambers or microfluidic chips wherein the depth of the liquid medium in the sample compartment is limited to 10 μm or less. Here we report an innovative and simple way to fabricate plastic devices with infrared transparent view-ports enabling infrared spectromicroscopy of living biological samples; therefore the device is named "IR-Live". Advantages of this approach include lower production costs, a minimal need to access a micro-fabrication facility, and unlimited mass or waste exchange for the living samples surrounding the view-port area. We demonstrate that the low-cost IR-Live in combination with microfluidic perfusion techniques enables long term (>60 h) cell culture, which broadens the capab...
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