Masson trichrome, a simple histological stain, was primarily used to distinguish collagen fibres and to detect collagen synthesis [31 (link), 32 (link)]. To determine whether the cutting plane in the superficial/deep DF and control groups was newly formed collagen deposition, masson trichrome staining was performed as previously described with some modifications [31 (link)]. Under 100 magnification, the cutting plane in each group was observed using a Zeiss microscope (Carl Zeiss, Inc., Germany) and was captured with a digital still camera (Nikon Instruments Inc., NY, U.S.A.).
Digital still camera
Manufactured by Nikon
Sourced in Germany
The Digital still camera is a device that captures and records photographic images in a digital format. It is designed to convert light into electrical signals, which are then processed and stored as digital data. The core function of the Digital still camera is to enable users to take high-quality photographs that can be easily shared, edited, and stored electronically.
Automatically generated - may contain errors
Lab products found in correlation
Inverted fluorescent microscope, by Nikon (1 mentions)
Anti cd19, by Wuhan Servicebio Technology (1 mentions)
Anti cd4, by Wuhan Servicebio Technology (1 mentions)
Anti il 17a, by Proteintech (1 mentions)
Anti ahr, by Abcam (1 mentions)
Ab5694, by Abcam (1 mentions)
Image pro plus, by Media Cybernetics (1 mentions)
4 protocols using digital still camera
1
Masson Trichrome Staining for Collagen Analysis
2
Dual Immunofluorescence Localization of Immune Markers
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Dual immunofluorescence was performed to colocalize CD4, CD19, IL-17A and IL-22 to AhR. Briefly, optimum cutting temperature embedded skin tissues were cut into 5-μm sections and boiled in ethylenediaminetetraacetic acid buffer (pH, 9.0) for antigen retrieval. The sections were incubated with the primary antibody overnight at 4°C. The following primary antibodies (mouse antihuman) were used: anti-CD4 (Servicebio, Wuhan, China, 1:100), anti-CD19 (Servicebio, Wuhan, China, 1:100), anti-IL-17A (Proteintech, Beijing, China, 1:200), anti-IL-22 (Bioss, Beijing, China, 1:100) and anti-AhR (Abcam, Cambridge, US). Then, the sections were incubated with corresponding fluorescence secondary antibody and counterstained using 4’,6-diamidino-2- phenylindole. The sections were observed under an inverted fluorescent microscope (Nikon, Tokyo, Japan) at original magnification ×200 under appropriate absorption/emission wavelengths (Alexa Fluor, 594 nm or 488 nm). The images were captured using a Nikon Digital Still Camera and analyzed by 2 independent investigators. Immunostained cells in the dermis were counted, and results were expressed as the number of positive cells/high power field (HPF). Interobserver variations for the counts were <10%.
3
Immunohistochemical Analysis of alpha-SMA Expression
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For the location of alpha smooth muscle actin (a-SMA), one of the key profibrotic markers that stimulate the development of fibrotic conditions [33 (link)–37 (link)], immunohistochemical staining was performed as previously described with some modifications [23 , 38 (link)]. Briefly, sections (4 μm) were deparaffinized with xylene and rehydrated and incubated with 3% H2O2 for 20 min to eliminate endogenous peroxidase activity. The sections were then incubated overnight at 4°C with a 1:1500 dilution of primary rabbit anti-a-SMA antibody (ab5694; Abcam, Cambridge, MA, U.S.A.), which was detected with a GTVision TM Ш Detection System/Mo&Rb (GK500705, Gene Tech Co., Ltd., Shanghai, China), as described by the manufacturer's instructions. The normal skin sample stained with primary antibody and the wounded skin sample stained with PBS served as controls. At 25, and 200 magnification, the location of a-SMA in each group was carefully examined using a Zeiss microscope (Carl Zeiss, Inc., Germany) and was captured with a digital still camera (Nikon Instruments Inc., NY, U.S.A.).
4
Choroidal Neovascularization Quantification
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5,7,9,11,14,17,19 or 21days after photocoagulation, the mice were perfused with 1 ml of PBS containing 50 mg/ml of fluorescein-labeled dextran (2×106 average molecular weight; Sigma-Aldrich, St. Louis, MO, USA) from the left ventricular, and euthanized 5 minutes later. Eyes were then enucleated and fixed in 4% formalin for 5 hours. Choroidal membranes were carefully dissected, flat-mounted, and examined by fluorescence microscopy. Images were captured with a digital still camera (Nikon Instruments Inc., New York, NY). Image analysis software (Image-Pro Plus; Media Cybernetics, Silver Spring, MD, USA) was used to measure the total area of CNV at each rupture site, with the investigator masked with respect to the experimental groups.
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