Characterization of thin section (egg SK1-5)
Thin sections of the eggshell were studied using transmitted light microscopy, X-ray radiography and X-ray diffraction.
For transmitted light microscopy, we used a Leica DMR XP microscope. Images were recorded with Leica DC 300 digital camera and the software Photostudio 5.5 (Arcsoft, Fremont, USA). Each frame was recorded twice, using polarized transmitted light and analysed polarized transmitted light.
The X-ray radiography of the thin section was performed at the ID19 beamline of the ESRF, using a 19.6 keV pink beam, a sCMOS pco.edge 5.5 camera, and an optic setup producing radiographs with an isotropic pixel size of 0.7 microns. Each radiograph was obtained from 1.2 seconds of exposure. The mesh of radiographs was performed at a sample-detector distance of 200 mm to observe phase contrast edge enhancement effect. We performed single-distance phase retrieval on the radiographs using the ANKA phase plug-in [17] of ImageJ [18]. The delta-beta value was set to 169. The radiographs obtain by phase-retrieval were stitched together using the Grid/Collection Stitching plug-in [19] of Fiji [20].
The diffraction experiment was performed on the ID19 beamline of the ESRF. We used a focused beam (Be lenses) and bended double Laue crystals to produced a monochromatic beam of 30 keV, cropped to obtain a pencil-beam of 50 x 50 μm. The images were recorded using a FReLoN 2k camera with a binning factor of 2, and an optic system producing images with an isotropic pixel size of 40 μm. The thin section was positioned perpendicular to the beam, and we analysed a 2.2 x 2.2 mm portion (S2 Fig). This portion was characterized step-by-step, recording independent diffraction pattern at individual points of the whole portion (89 x 89 points). Each diffraction pattern was obtained from 2 seconds of exposure and the thin section was displaced by 25 μm, either horizontally or vertically, to create a diffraction mapping. It resulted in 7921 diffraction patterns. Because for each recorded image, only a few crystals were oriented in Bragg conditions, we produced maximum intensity projections of several diffraction images representing various area of the studied portion. From these projection images, we performed 360° azimuthal integration, resulting in a plot of intensity of the diffractions circles (represented as peaks) versus distance to the centre of the image, in pixel. As the wavelength of the incoming X-ray beam was quite unusual (0.413 Å compared to more common 1.54 Å obtain from Cu Kα) it was decided to convert the distance to the centre values into d-spacing values rather than 2θ angles. The resulting diffraction patterns were compared with standards from the literature to identify dominant mineral phases.
คุณสมบัติของบางส่วน (ไข่ SK1-5)ส่วนที่บางของเปลือกไข่ถูกศึกษาใช้ microscopy นำส่งแสง radiography เอกซเรย์ และการเลี้ยวเบนเอ็กซ์เรย์นำส่งแสง microscopy เราใช้กล้องจุลทรรศน์ไล XP ทรัพยากรธรณี ได้รับการบันทึกภาพ ด้วยกล้องดิจิตอลไล DC 300 และซอฟต์แวร์ Photostudio 5.5 (Arcsoft, Fremont สหรัฐอเมริกา) เฟรมแต่ละถูกบันทึกสอง ใช้แสงโพลาไรซ์นำส่งและ analysed ขั้วนำส่งแสงทำ radiography เอกซเรย์ส่วนบางที่ beamline ID19 ของ ESRF ใช้แสงสีชมพู keV 19.6 กล้อง sCMOS pco.edge 5.5 และการติดตั้งใยแก้วนำแสงที่ผลิต radiographs ด้วยขนาดพิกเซล isotropic 0.7 microns Radiograph แต่ละได้รับจาก 1.2 วินาทีของการสัมผัส ตาข่ายของ radiographs ที่ดำเนินการในตัวอย่างจับสังเกตลักษณะปรับระยะความคมชัดขอบ 200 mm เราดำเนินการเรียกระยะห่างเดียวกับ radiographs ใช้ระยะ ANKA ปลั๊กอิน [17] ของ ImageJ [18] ค่าเบต้าเดลต้าถูกตั้งค่าให้ 169 Radiographs ได้รับ โดยเรียกระยะถูกเย็บเข้าด้วยกันโดยใช้กริด/ชุดเย็บปลั๊ก [19] [20] ฟิจิThe diffraction experiment was performed on the ID19 beamline of the ESRF. We used a focused beam (Be lenses) and bended double Laue crystals to produced a monochromatic beam of 30 keV, cropped to obtain a pencil-beam of 50 x 50 μm. The images were recorded using a FReLoN 2k camera with a binning factor of 2, and an optic system producing images with an isotropic pixel size of 40 μm. The thin section was positioned perpendicular to the beam, and we analysed a 2.2 x 2.2 mm portion (S2 Fig). This portion was characterized step-by-step, recording independent diffraction pattern at individual points of the whole portion (89 x 89 points). Each diffraction pattern was obtained from 2 seconds of exposure and the thin section was displaced by 25 μm, either horizontally or vertically, to create a diffraction mapping. It resulted in 7921 diffraction patterns. Because for each recorded image, only a few crystals were oriented in Bragg conditions, we produced maximum intensity projections of several diffraction images representing various area of the studied portion. From these projection images, we performed 360° azimuthal integration, resulting in a plot of intensity of the diffractions circles (represented as peaks) versus distance to the centre of the image, in pixel. As the wavelength of the incoming X-ray beam was quite unusual (0.413 Å compared to more common 1.54 Å obtain from Cu Kα) it was decided to convert the distance to the centre values into d-spacing values rather than 2θ angles. The resulting diffraction patterns were compared with standards from the literature to identify dominant mineral phases.
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