Fossil specimens were photographed with a Keyence VHX 6000 and Leica M205AF stereomicroscope. In order to observe the structures buried within the rock matrix and to produce three-dimensional models of the preserved morphology, micro-CT imaging and three-dimensional computer rendering techniques were applied. The best outcome is from specimens YKLP 16260 and CFM 00047. Specimen YKLP 16260 was first scanned with a GE Phoenix Nanotom cone beam scanner at the Bavarian State Collection of Zoology, Bavarian Natural History Collections, München, Germany, to detect the signal/noise ratio, and then with a Zeiss Xradia 520 Versa X-ray microscope at Yunnan Key Laboratory for Palaeobiology to obtain images with higher resolutions. For the Xradia 520 Versa scanning, the energy and the resolution were set at 60 kv/5w and 15.25 µm for slab a, and at 60 kv/5w and 15.89 µm, 50 kv/4w and 8.48 µm for slab b (scanned twice), to obtain a higher resolution of the anterior part of the body. Specimen CFM 0047 was scanned with a Zeiss Xradia 520 Versa X-ray microscope with the above two parameters set at 70 kv/6w and 5.41 µm for overview scanning, and at 60 kv/5w and 11.6 µm for small-field, detailed scanning.
Xradia 520 versa x ray microscope
Manufactured by Zeiss
Sourced in Germany, China
The Xradia 520 Versa X-ray microscope is a non-destructive imaging system designed for high-resolution 3D visualization and analysis. It utilizes advanced X-ray technology to capture detailed internal structures and compositions of a wide range of samples, including materials, devices, and biological specimens.
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Lab products found in correlation
Ccd camera, by Oxford Instruments (1 mentions)
Vhx 6000, by Leica camera (1 mentions)
Phoenix nanotom, by GE Healthcare (1 mentions)
Em cpd300, by Leica camera (1 mentions)
Smz1500 stereoscopic microscope, by Nikon (1 mentions)
2 ml tube, by Eppendorf (1 mentions)
Eos 760d, by Canon (1 mentions)
Focus 6, by Helicon (1 mentions)
Smz1500, by Nikon (1 mentions)
1.5 ml tube, by Eppendorf (1 mentions)
Xmreconstructor, by Zeiss (1 mentions)
14 protocols using xradia 520 versa x ray microscope
1
Micro-CT Imaging and 3D Modeling of Fossil Specimens
2
3D Imaging of Subcutaneous Tissue
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The 3D imaging of SC was performed using the ZEISS Xradia 520 Versa X-ray Microscope [Carl Zeiss (Shanghai) Co. Ltd., Shanghai, China]. The specimens were stained with 1% osmium acid solution and dried with the critical point drying method. Before imaging, the specimens were mounted on the sample holder with an aluminum tube as adapter and rotated horizontally by 360°, pausing at discrete angles to collect 2D projection images, which were then combined together to produce a 3D reconstruction of the specimen’s volume dataset. Scanning energy is 40 to 60 kV, and scanning resolution is from 0.5 to 2.0 μm/voxel size. The low resolutions are for the large filed of sample scanning, and fine resolutions are for the local high-resolution scanning. Each 3D dataset includes about 1000 virtual slices. The volume of 3D datasets is about 2 GB in CCD (Charge-coupled Device) bin2 mode. Dragonfly software from Object Research Systems (ORS) Inc. is hired for dataset processing. The 3D internal structure of SC is presented after segmentation and 3D rendering.
3
Imaging Implanted Rat Sciatic Nerves
4
Characterizing Porosity in Additive Manufacturing
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Phase determination was carried out by X-ray diffraction (XRD) using a C8 Discover (Bruker Corporation, Billerica, MA, USA) with CuKα radiation (λ = 1.540562 Å). The samples were scanned continuously from 15° to 88° in 2θ with a step size of 0.01°.
The porosity of the as-cast samples compared with the AM-generated parts with different process parameters was investigated with an Xradia 520 Versa X-ray microscope (Carl Zeiss AG, Oberkochen, Germany). The images were taken with a CCD camera (Andor Technology Ltd., Belfast, Northern Ireland, UK) and lenses at 4 times magnification. For the as-cast samples, a spatial resolution of 2.4 µm, an acceleration voltage of 70 kV, a power of 6 W and an exposure time of 3.2 s were used. The tomographic images of the parts generated with AM were implemented with spatial resolutions of 5 (samples for the compression test) and 6 µm (samples for bending tests), an acceleration voltage of 80 kV, a power of 7 W and exposure times of 5 (samples for the compression test) and 8 s (samples for bending tests).
The porosity of the as-cast samples compared with the AM-generated parts with different process parameters was investigated with an Xradia 520 Versa X-ray microscope (Carl Zeiss AG, Oberkochen, Germany). The images were taken with a CCD camera (Andor Technology Ltd., Belfast, Northern Ireland, UK) and lenses at 4 times magnification. For the as-cast samples, a spatial resolution of 2.4 µm, an acceleration voltage of 70 kV, a power of 6 W and an exposure time of 3.2 s were used. The tomographic images of the parts generated with AM were implemented with spatial resolutions of 5 (samples for the compression test) and 6 µm (samples for bending tests), an acceleration voltage of 80 kV, a power of 7 W and exposure times of 5 (samples for the compression test) and 8 s (samples for bending tests).
5
X-ray Microscopy of Eragrostis tef Spikelets
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Sample preparation and XRM imaging methods were adapted from Duncan et al. (2022) (link). A combination fixation-contrast solution, ethanolic phosphotungstic acid (ePTA), was prepared at 1% PTA (w/v) in 70% ethanol (v/v). Live E. tef spikelets were transferred to 10 mL of ePTA in 20 mL glass sample vials and placed on a lab rocker at room temperature (RT) for at least 21 d before imaging, and the ePTA solution was exchanged at 7 and 14 d. For XRM imaging, a single fixed and contrast-enhanced spikelet was rinsed for 10 min in distilled water, then transferred to a 0.2 mL flat-cap PCR tube filled with melted low melting point agarose at 2% (w/v) in distilled water. When the agarose had solidified, the sample tube was mounted on the end of a wooden applicator stick using a 2-stage 5-min epoxy resin and placed in the XRM sample holder. Imaging was done on a ZEISS Xradia 520 Versa X-ray microscope using a 20× objective lens with the following instrument settings: 60 kV, 5 W, 83 μA, binning = 2, exposure = 7 s, 4,001 projections, geometric magnification = 1.8× for a voxel resolution of 0.7 μm. Scans were automatically reconstructed using ZEISS Reconstructor software, and image analysis used ORS Dragonfly for noncommercial users. Lemma width was estimated using the formula area/perimeter*2 measured by Dragonfly software (Object Research Systems, Montréal, Canada).
6
Critical Point Drying for X-Ray Microscopy
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TQ and nal8 seedlings were grown in an illuminated chamber for 7 days after germination. Samples were collected and fixed in FAA (50% ethanol, 5% glacial acetic acid, 5% formaldehyde) for 12 h at 4 °C. After the tissue was dehydrated in a graded ethanol series, the samples were thoroughly desiccated in an automated Critical Point Dryer (Leica EM CPD300). Samples were observed with a Zeiss Xradia 520 Versa X-Ray Microscope. XM 3DViewer was used to generate the visual image slices.
7
Cenomanian-Albian Amber Specimen Analysis
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The specimen was collected from Noije Bum 2001 Summit Site, Hukawng Valley, Kachin, Myanmar (26°20′N, 96°36′E) (Fig. 1 ). Paleontological studies indicate that the specimen belongs to the earliest Cenomanian-latest Albian, Early Cretaceous (98.79 Ma)34 (link), which is generally agreed on by Poinar et al.28 (link) and Xing et al.49 . Two parallel planes were made on the amber sample before observations. Observations and photographs were made with a Nikon SMZ1500 stereoscopic microscope at the Nanjing Institute of Geology and Palaeontology, Nanjing, China. Micro-CT was performed using a Zeiss Xradia 520 versa X-ray microscope at the Nanjing Institute of Geology and Palaeontology, Nanjing, China. The 3D reconstruction and virtual sections were generated using VGStudio MAX 3.0. All figures were organized for publication using Photoshop 7.0.
8
High-Resolution X-ray Tomography Protocol
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The measurement was done using a ZEISS Xradia 520 Versa X-ray microscope with a Flat Panel (FPX) detector. The polychromatic micro-focus source was operated at 80 kV, 7 W and with a low energy filter (LE2) to optimise transmission. In total, 3201 radiographs were acquired over 360 sample rotation with an exposure time of 1 s per radiograph, resulting in a total scan length of 1 h 30 min. The transmission images were reconstructed using the commercial software from ZEISS to an isotropic voxel size of .
9
Quantifying Quadriceps Muscle Mineralization
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Animals were harvested at 8 weeks of age. The left quadriceps muscle was fixed in 10% formalin for 48 h, then to transferred to 70% ethanol prior to scanning. Quadriceps were placed within a 2-ml tube (Eppendorf, Stevenage, United Kingdom) and supported by a polyurethane foam saturated in 70% ethanol. Muscles were imaged using a Zeiss Xradia 520 Versa X-ray microscope (Zeiss) operating at an energy of 50 kV, a power of 4 W, and a tube current of 80 μA with a Zeiss LE1 filter was positioned directly after the X-ray source. The sample distance to the x-ray source was 200mm, and the sample distance to detector was 225mm, A total of 1601 X-ray projection images were collected over 360° at equal intervals with an exposure time for each projection of 3 s. The projections were reconstructed using the manufacturer’s integrated software, which uses a filtered back projection reconstruction algorithm. TXRM files were imported into FiJi with the plugin to read XRM files and export Z-stack images (Schindelin et al., 2012 (link); Metscher 2020 ). Mineralised areas within exported z-stack images (pixel size ranging from 32.25 to 34.45 µM depending upon sample) were quantified using the threshold function in ImageJ and used to calculate the volume.
10
Fossil Fruit Imaging and Analysis
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Fossil fruits were cleaned with an ultrasonic cleaner (JieMeng JP–880) at a frequency of 42 kHz and photographed using a Nikon SMZ25 stereo microscope at the Museum of Biology, Sun Yat-sen University (Guangzhou, China). Extant fruits were photographed by a Canon EOS 760D digital camera. DigiCamControl-Free Windows DSLR camera controlling solution,54 Helicon Focus 6.6.1 (Helicon Soft Ltd., Kharkov, Ukraine) and Photoshop 2020 (Adobe, San Jose, California, USA) were used for image processing. Microstructure was examined using a thermal field emission environmental scanning electron microscope (Quanta 400) at the test center, Sun Yat-sen University (Guangzhou, China). The internal three-dimensional structure of fruit was observed by Zeiss Xradia 520 Versa X-ray microscope (CT) housed at the University of Science and Technology of China (Hefei, China) and image data were processed using Dragonfly software.
The assumption that Cenozoic extinct plants and their nearest living relatives (NLRs) have similar environmental requirements, is the base of the NLR method. This method was used to estimate the paleoclimatic and paleoecologic conditions under which the studied fossil taxon lived by identifying the NLRs of fossil species and the environmental characteristics predominating in the regions of their modern natural distribution.55
The assumption that Cenozoic extinct plants and their nearest living relatives (NLRs) have similar environmental requirements, is the base of the NLR method. This method was used to estimate the paleoclimatic and paleoecologic conditions under which the studied fossil taxon lived by identifying the NLRs of fossil species and the environmental characteristics predominating in the regions of their modern natural distribution.55
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