Photographs were taken before the first laser treatment (baseline, week 0), before the further laser treatments (week 2, 4 and 6) and at the final assessment (week 8) (Figure 1 ). All photographs were taken by one photographer with a Nikon D200 camera, under identical conditions of illumination and patient positioning.
D200 camera
Manufactured by Nikon
Sourced in Japan, United States
The Nikon D200 is a digital single-lens reflex (DSLR) camera. It features a 10.2-megapixel CCD image sensor, 5-area autofocus system, and 2.5-inch LCD display. The camera is capable of capturing high-quality still images and supports a range of interchangeable lenses.
Automatically generated - may contain errors
Lab products found in correlation
Af micro nikkor 60 mm lens, by Nikon (1 mentions)
C57bl 6j, by Jackson ImmunoResearch (1 mentions)
Xylol, by Carl Roth (1 mentions)
Hcl 1n, by Merck Group (1 mentions)
Metavue, by Universal Imaging (1 mentions)
Imagej, by Universal Imaging (1 mentions)
Dissection microscope, by Olympus (1 mentions)
Betadine, by Sanofi (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
8 protocols using d200 camera
1
Standardized Photographic Documentation of Laser Treatments
2
Quantitative Immunoblot Analysis of IRAK1 and IKKβ
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RAW264.7 cells were transfected with either SF3a1-specific siRNA or control non-targeting siRNA as described above. Following the siRNA treatment, cells were lysed on ice in RIPA buffer supplemented with protease inhibitors. Lysates were centrifuged at 12,000 RPM for 15 minutes at 4°C, protein concentration of the supernatant was assessed by BCA Assay (Pierce), and samples were boiled in SDS-loading buffer. Samples were separated on 10% SDS-polyacrylamide gels and transferred to nitrocellulose. The membranes were blocked for 2 hours at room temperature in TBS-T containing 5% non-fat milk, incubated overnight at 4°C with primary antibodies (1:1000) in TBS-T plus 5% BSA (rabbit-anti-IRAK1 and rabbit-anti-IKKβ antisera were from Cell Signaling Technology; mouse-anti-β-actin antiserum was from Millipore), washed in TBS-T, then incubated with HRP-conjugated secondary antibodies (1:1000) for 1 hour at room temperature. The membrane was then washed, treated with ECL Substrate (Pierce), and fluorescence was captured by autoradiography. Images of the films were captured with a Nikon D200 camera. Bands were quantified using Image J [142 (link)] and subsequently analyzed for significant differences in Graphpad Prism 5 using t tests (p<0.05).
3
Fossil Pinites Collection and Imaging
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All specimens used in this study (S1 Table ) were borrowed from the Herbarium of the University of Tokyo (TI) and the Fossil collections of the Osaka Museum of Natural History (OSA; for details on these herbaria including contact information, see Index Herbariorum [35 ]). No other specimens were used in this study. Specimens were photographed using a D200 camera (Nikon, Tokyo, Japan) with an AF MICRO NIKKOR 60 mm lens (Nikon) under fluorescent illumination.
The holotype of Pinites fujiii stored in TI consists of a female cone, a replica of the cone, and four microscope slides mounting sectioned parts of the cone. No specimen number is assigned to the holotype, while it is registered as “holotype of Pinites fujiii”. The holotype was collected in Seto-shi, Aichi Pref., Japan (Fig 1 ), from the Seto Formation, but the exact locality is not available.
Other specimens, including Miki’s [30 ] specimens, are stored in OSA F. These specimens were slightly compressed mummifications collected from the Seto or Tokiguchi Formations (Fig 1 ).
The holotype of Pinites fujiii stored in TI consists of a female cone, a replica of the cone, and four microscope slides mounting sectioned parts of the cone. No specimen number is assigned to the holotype, while it is registered as “holotype of Pinites fujiii”. The holotype was collected in Seto-shi, Aichi Pref., Japan (
Other specimens, including Miki’s [30 ] specimens, are stored in OSA F. These specimens were slightly compressed mummifications collected from the Seto or Tokiguchi Formations (
4
Corneal Wound Healing in Mice
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Corneal wounding experiments in mice were conducted in compliance with the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research. The protocol was approved by the UIC Animal Care and Use Committee. Twelve to 19-week-old C57BL/6J (Jackson Laboratory, Bar Harbor, ME, USA) mice were anesthetized with intraperitoneal injection of ketamine (100 mg/kg) and xylazine (5 mg/kg). After applying two drops of topical 0.5% proparacaine, a 2.0-mm area of the central epithelium was demarcated using a 2-mm disposable biopsy punch and removed by an AlgerBrush II (The Alger Company, Lago Vista, TX, USA). In the treatment (n = 7) or the control (n = 7) group, Hst5 (80 μM), SHRGY (80 μM), or SP1 (80 μM) was applied to the cornea three times a day. At 0, 18, and 24 h, the corneas were stained with fluorescein (FUL-GLO fluorescein sodium ophthalmic strips, Akorn, Lake Forest, IL, USA) and imaged using a Nikon FS-2 photo-slit lamp with a Nikon D200 camera (Nikon, Melville, NY, USA). At each indicated time point, the remaining wound areas were measured using ImageJ software and compared with the baseline wound area for each mouse, and the percentage of the remaining wound area was calculated for each time point.
5
Mammary Gland Whole Mount Analysis
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Mammary gland whole mount analysis was carried out as described by Kleinberg et al. [23] (link). All chemicals came from Merck (Darmstadt, Germany), unless otherwise stated. After harvesting, tissues were fixed in ice-cold 4% paraformaldehyde for 2 h. Mammary glands were stored in 70% ethanol until further analysis. In short, following 3 washes with aceton for 30 min, mammary glands were rehydrated in 100% and 95% ethanol (30 min each). The glands were stained for 1 h with filtered hematoxylin, followed by thorough rinsing with tap water. Background staining was minimized by incubation (3×35 min) in a solution of 150 ml 100% ethanol, 150 ml aqua dest. and 7.5 ml HCl 1N (Sigma-Aldrich Chemie GmbH, Munich, Germany). Subsequently tissues were dehydrated in 70%, 95% and 100% ethanol (2×30 min for each step), followed by an incubation in Xylol (Roth, Karlsruhe, Germany) overnight. For long term storage mammary gland whole mounts were placed in methylsalicylate. Digital images of squash preparations were taken by a transmitted-light microscope stand mounted with a NIKON D200 camera (Nikon GmbH, Düsseldorf, Germany) at a distance of 60 cm all in one day. Picture analysis was performed using MetaVue software (MetaVue, Universal Imaging Corp., Downingtown, PA, USA) and ImageJ [34] (link). Pixel size was 11.125 µmx11.125 µm and pixel area was 123.766 µm2.
6
Standardized Facial Photography Protocol
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Photographs of the patients' faces lateral of the right and left eyes were taken under standardized conditions at Wk 0, Wk 4, Wk 8, and Wk 12. All photographs were taken with participants looking straight ahead under constant lighting conditions using the same camera, camera settings, and camera placement. A Nikon D200 camera (Nikon, Tokyo, Japan) was positioned on an SVK 35D tripod (Slik, Saitama, Japan) to the right of the participant's face and perpendicular to the area to be photographed. Simultaneously, an Olympus twin T28 Macroflash unit (Olympus, Shinjuku, Japan) was positioned with one flash head directly above the lens pointed at the participant and the other directed away. Exposure was controlled by adjusting the lens aperture with the flash set on manual at full output.
7
Corneal Wound Healing Assay in Mice
8
A Head Biopsy Model for Wound Healing Assessment
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A head biopsy model was employed, as recommended for assessment of growth factors in wound healing [18 (link),19 ]. Briefly, five female BALB/c mice per group (rTRX, PBS and rOv-GRN-1) were anesthetized (intraperitoneal xylazine 16 mg kg-1; ketamine 80 mg kg-1), after which the crown of the head was shaved with an electric razor. Mice were anesthetized three days later and the surgical site was sterilized with 70% ethanol wipes. A skin-deep wound of 5 mm in diameter was inflicted on the crown of the head using biopsy punch (Zivic instruments). The lesion was rinsed with antiseptic (Betadine, Sanofi), after which 56 pMoles of rOv-GRN-1, rTRX or PBS suspended in 1.5% methyl cellulose (Sigma) in 50 μl was applied. Thereafter, the lesion was covered with Elastoplast Spray Plaster (Beiersdorf). Progress of the wound, and wound closure, was documented with photographs taken at cumulative 1.6× magnification using a dissection microscope (Olympus) fitted with a Nikon D200 camera, each day for five days. Wound closure was ascertained in an unblinded fashion by comparison of the surface area of the lesion with the size as documented immediately after the wound was inflicted, with the assistance of ImageJ software.
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