After deparaffinization and processing for antigen retrieval, the blank colon sections were incubated overnight at room temperature with anti-rabbit ZO-1 antibody (1:500, catalog no. GB11195; Servicebio, China) and Occludin antibody (1:200, catalog no. GB11149; Servicebio) separately. Sections were then incubated with anti-rabbit IgG secondary antibody (KPL, MA, USA) for 50 min at room temperature. After washing with PBS, the DAB Envision kit (catalog no. K5007; Dako, Copenhagen, Denmark) was used to develop color; ZO-1 and Occludin appear brown, while nuclei are blue. Images of each colon section were obtained in tripartite by experienced staff who were blind to the experiment under ×200 magnification using a Leica DMRBE microscope and were analyzed using Image Pro Plus 6.0, according to the method previously described [47 (link)]. The integrated optical density (IOD) values were log10 transformed.
Dmrbe microscope
Manufactured by Leica
Sourced in Germany
The DMRBE microscope is a high-performance research microscope designed for advanced microscopy applications. It features a modular design, allowing for customization and adaptation to various experimental requirements. The DMRBE provides reliable and precise optical performance, enabling users to capture detailed images and conduct thorough analyses.
Automatically generated - may contain errors
Lab products found in correlation
Axiovision rel 4, by Zeiss (3 mentions)
Paraplast plus, by Merck Group (3 mentions)
Tcs sp2 scanning head, by Leica (3 mentions)
Application suite v3, by Leica (3 mentions)
Dfc420c camera, by Leica (3 mentions)
Envision dab kit, by Agilent Technologies (2 mentions)
Gb11149, by Wuhan Servicebio Technology (2 mentions)
Lsm 780, by Zeiss (2 mentions)
Axiocam hr, by Zeiss (2 mentions)
Dcf480, by Leica (2 mentions)
Biotinylated swine anti rabbit immunoglobulin, by Agilent Technologies (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Image pro plus 6, by Leica (1 mentions)
Avidin biotin blocking kit, by Vector Laboratories (1 mentions)
Periodic acid solution, by Merck Group (1 mentions)
Fisher chemical permount mounting medium, by Thermo Fisher Scientific (1 mentions)
Photo flo, by Kodak (1 mentions)
Dfc320 camera, by Leica (1 mentions)
Anti acetylated α tubulin, by Merck Group (1 mentions)
Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions)
61 protocols using dmrbe microscope
1
Immunostaining of Tight Junction Proteins
2
Stem Cell Wall Lignification Imaging
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Hand-cut stem sections from the first internode (1 cm below first node below spikelet) of plants 39 days after planting were made with a Teflon-coated razor blade (GEM, Ted Pella). Sections were stained 15 min in acid phloroglucinol (2% (w/v) phloroglucinol/20% (v/v) ethanol/2.4 m HCl), mounted on microscope slides, and then photographed using a Leica DMRBE microscope with brightfield illumination. Care was taken to stain each section the same duration of time before imaging. Mäule method analysis was performed as in Bouvier d'Yvoire et al. (2013) (link).
3
Fluorescent Immunohistochemical Imaging Workflow
4
Colony Formation Assay for MGC803 Cells
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MGC803 and MGC803‐resistant cells were trypsinized and resuspended in growth medium, and then seeded into four‐six‐well plates (200 cells in each well). The cells were then treated with DMSO and 10058‐F4, respectively, (four groups: MGC803/DMSO, MGC803/10058‐F4, MGC803‐resistant/DMSO, and MGC803‐resistant/10058‐F4). Next, when the cultured cells grew and formed the visible cell colonies, the cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. The number of colonies was counted under a DM‐RBE microscope (Leica, Heidelberg, Germany). Each experiment was performed in triplicate.
5
Immunohistochemical Staining of Spleen Tissue
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Snap frozen spleen sections (5 μm) were fixed (chloroform/acetone, 1:1, 4 min) and treated with levamisole to ablate tissue alkaline phosphatase activity. Nonspecific binding was blocked using an avidin-biotin-blocking kit (VectorLaboratories, Burlingame, CA, USA), and 2% normal serum derived from the same species as the secondary antibodies. Tissues were incubated with the primary antibodies, washed, exposed to biotinylated secondary antibodies and alkaline phosphatase conjugated avidin-biotin solution and were counterstained with hematoxilin. Images were taken using a Leica DMRBE microscope.
6
Immunofluorescent Staining on Glass Slides
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Cells on glass-slides were fixed, permeabilized, blocked, incubated with primary antibody, fluorochrome-conjugated secondary antibody, blocked, incubated with second, dye-labeled primary antibody and washed. Slides were mounted in Elvanol. Digitized images were generated using a Leica DMRBE microscope.
7
Immunohistochemistry of Frozen Tissue
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Shock-frozen tissue sections (8 μm) were fixed, incubated with antibodies, washed, exposed to biotinylated secondary antibodies and alkaline phosphatase-conjugated avidin-biotin solution. Sections were counter-stained with hematoxilin. Digitized images were generated using a Leica DMRBE microscope.
8
Quantification of Colonic Goblet Cells
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The 5-μm-thick sections were fixed with 10% formalin followed by hydration in water. The tissue was stained with periodic acid solution (Sigma) for 5 min, Schiff's reagent for 15 min, and hematoxylin solution for 90 sec. Samples were dehydrated in increasing concentrations of ethanol (70%, 90%, 95%, and 100%) followed by incubation in xylene and finally sealed with a coverslip using Fisher Chemical™ Permount™ Mounting Medium (Fisher Scientific). Goblet cells were observed and counted in individual well-preserved colonic crypts using a Leica DMRBE microscope.
9
Immunohistochemical Tissue Imaging
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Snap frozen sections (5 μm) were fixed, incubated with antibodies, washed, exposed to biotinylated secondary antibodies and alkaline phosphatase conjugated avidin-biotin solution. Sections were counter-stained with H&E. Digitized images were generated using a Leica DMRBE microscope.
10
Imaging Worm Morphology on Microscope
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Images were taken on a Leica DMRBE microscope using a 10×/0.30 PL FLUOTAR objective. A QIClick (QIMAGING) camera captured images using Q-Capture software. Scale bars in the whole worm images represent 50 μm.
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