ECMs were synthesised on glass coverslips as described before. Cells were then plated on the ECM for the duration indicated elsewhere. Fixing of the cells was achieved using 3% paraformaldehyde solution. Permeabilisation of the cells was done using 0.1% Triton X-100 (Sigma Aldrich Chemie, Steinheim, Germany) in PBS. Blocking of cells was done using 1% BSA for 1 h at room temperature and then incubated with various primary antibodies overnight at 4 °C. Cells were then washed three times with PBS. Secondary antibodies were conjugated to FITC and Fluor 488. DAPI was added in order to visualise the nucleus. Fluorescence was observed using a Zeiss Inverted Microscope with a 20× objective. Acquisition of images was achieved using the CellSens Imaging System (Olympus, Tokyo, Japan). Proteoglycan composition within the ECMs was detected by staining the ECMs with 1% Alcian blue after preparation using a standard protocol. Briefly ECM synthesis and preparation was done on coverslips. ECMs were incubated with 1% Alcian Blue solution for 20 min and washed three times. Images were observed and photographed using a light microscope (Olympus CKX41 with SC30 camera). Images were taken at 100× magnification.
Cellsens imaging system
Manufactured by Olympus
Sourced in Japan
The CellSens Imaging System is a microscope imaging solution designed for cell biology and life science research. It offers advanced imaging capabilities for capturing and analyzing cellular samples. The system provides high-quality image acquisition and processing functionalities to support various microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Inverted microscope, by Zeiss (2 mentions)
Sc30 camera, by Olympus (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Ckx41, by Olympus (1 mentions)
Bx46 microscope, by Olympus (1 mentions)
Eos 6d digital camera, by Canon (1 mentions)
Citrate buffer, by Agilent Technologies (1 mentions)
3 3 diaminobenzidine chromogen, by Agilent Technologies (1 mentions)
Hrp conjugated goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Anti mouse cd31, by R&D Systems (1 mentions)
5 protocols using cellsens imaging system
1
Visualizing Cell-ECM Interactions
2
Histological Assessment of Implant Impact
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Histology was performed to assess the impact of placing the implant and electrical stimulation on tissue integrity. A bilateral craniotomy was performed in two animals prior to administering stimulation and two identical pieces of dura substitute were introduced in each animal, as shown in Supplementary Material Fig. S6 . The implant’s electrode made contact with one dura graft site (stimulation) and there was no contact with the other dura graft site (sham). The stimulation experiment was performed for at least 180 minutes. At the end of the stimulation experiment, the animal was placed in deep anesthesia by a ketamine-xylazine injection. The heart was exposed and a small cut was made in the left ventricle. A perfusion needle (15 ga) was inserted through the ventricle until it entered the aorta. A large incision was made in the right atrium to allow the blood to flow and drain. 200 mL of PBS was delivered followed by 200 mL of 4% paraformaldehyde. The flow was maintained at 20 mL/min. Rat brains used for H&E staining were embedded in paraffin wax following fixation and sectioned into 5 μm slices. Histology images were viewed with an Olympus BX46 microscope and selected images were captured using Standard Olympus cellSens imaging system. Whole brain slices were photographed at 12.5X and focus of ischemia at 100X.
3
Visualization of TB Lung Pathology
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Expression of CH25H and ADRP were detected in Mtb-infected mice, rabbits, marmosets, and clinical tuberculosis patients. The paraffin-embedded lung tissues of Mtb aerosol-infected mice, rabbits, and marmosets (under protocols LCIM-3, LCIM-4, and LCIM9) were obtained from the Tuberculosis Research Section of the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD.58 (link),59 (link) Paraffin-embedded clinical tuberculosis patient samples were provided by the Third People’s Hospital of Yichang and approved by the Ethics Committee of the Third People’s Hospital of Yichang. Five-micrometer serial sections were made from all tissues and stained with Ziehl-Neelsen (Z.N), anti-CH25H and anti-ADRP antibodies, and images were captured by using the Cellsens imaging system (Olympus Life Sciences).
4
Comparative Growth of P. oxalicum on Diverse Carbon Sources
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Spore suspensions of P. oxalicum strains were spread on solid plates supplemented with different carbon sources and cultured for 4 days at 28 °C. Carbon sources were raw cassava starch, soluble corn starch, wheat bran plus Avicel, PDA and glucose. Colony photographs were taken with an EOS-6D digital camera (Canon, Tokyo, Japan). A Cellsens Imaging system (Olympus, Tokyo, Japan) was also employed. All studies were performed in triplicate.
5
Immunohistochemistry of Vascular Markers
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Formalin‐fixed paraffin‐embedded tissue sections were dewaxed, and antigens were retrieved after 10 minutes of boiling in citrate buffer, pH 6 (Dako). Slides were stained with 10 μg/ml of biotinylated DVD antibodies for 1 hour at room temperature and were visualized with streptavidin–HRP complex using 3,3′‐diaminobenzidine chromogen (Dako). Rabbit anti–ICAM‐1 IgG (Abcam) was detected with HRP‐conjugated goat anti‐rabbit IgG (Jackson Immunotools). Mouse anti–von Willebrand factor (Dako) and mouse anti‐CD31 (R&D Systems) were used to identify human vascular endothelial cells, which were revealed with HRP‐conjugated goat anti‐mouse IgG (Santa Cruz Biotechnology). Sections were counterstained with hematoxylin, mounted with Depex mounting medium (Dako), and acquired with a CellSens imaging system (Olympus).
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