Neurological regeneration was tested by Nissl staining and Luxol fast blue staining [45 (link)] according to the instruction of manufacture. Rats were anesthetized by 3% pentobarbital sodium (42 mg/kg), injection and intracardially perfused with paraformaldehyde. The brain tissues were isolated and fixed in 4% paraformaldehyde for 24 hours, after which the hippocampus and cerebral cortex of the brain tissue were isolated. The brain tissues were dehydrated with 30% sucrose solution at 4°C for 3–5 days. After that, the brain tissues were preserved in opti-mum cutting temperature compound (OCT) for 6 hours at room temperature. Then the brain tissues were sliced into 8 and 20 μm thick sections at −20°C, respectively. Luxol fast blue (LFB) staining can demonstrated myelin. Briefly, 8 μm coronal brain slices were placed in LFB solution (Solarbio, China) at 60°C for 4 hours, then transferred to Lithium Carbonate for 30 seconds, obtained the image under bright field microscope (Leica, Germany). Nissl staining can demonstrate Nissl bodies. Briefly, 20 μm coronal brain slices were dehydrated in graded alcohol, staining by 0.1% cresyl violet (Solarbio, China), dehydrated in graded alcohol and xylene again, then transfer to Nissl Differentiation Solution (Solarbio, China) for 30 seconds, obtained the image under bright field microscope (Leica, Germany).
Bright field microscope
Manufactured by Leica camera
Sourced in Germany, United States
The Bright field microscope is a type of optical microscope that uses transmitted illumination to create an image of the sample. It is designed to provide a clear and detailed view of samples by illuminating them from below and allowing the light to pass through the specimen before reaching the objective lens. This microscope is commonly used in various scientific and research fields for the observation and analysis of biological and materials samples.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510 meta confocal microscope, by Zeiss (4 mentions)
Cm1850, by Leica camera (2 mentions)
Hematoxylin, by HiMedia (2 mentions)
Oil red o solution, by Merck Group (2 mentions)
Oro staining solution, by Sangon (2 mentions)
Crystal violet, by Merck Group (2 mentions)
Imagej, by Leica camera (2 mentions)
Alexa fluor conjugated secondary antibodies, by Santa Cruz Biotechnology (2 mentions)
Plan apochromat oil immersion objective, by Zeiss (2 mentions)
Zen software, by Zeiss (2 mentions)
A48282, by Thermo Fisher Scientific (2 mentions)
Optimal cutting temperature compound, by Sakura Finetek (2 mentions)
Superfrost plus slide, by Thermo Fisher Scientific (2 mentions)
Aec101 1kt, by Merck Group (1 mentions)
Primary antibodies against cd31, by Santa Cruz Biotechnology (1 mentions)
3 amino 9 ethylcarbazole, by Merck Group (1 mentions)
Biotinylated anti mouse igg, by Merck Group (1 mentions)
Extravidin peroxidase conjugate, by Merck Group (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
Anti periostin, by Abcam (1 mentions)
73 protocols using bright field microscope
1
Neurological Regeneration Assessment by Histological Staining
2
Senescence Evaluation in Cardiomyocytes
3
Colony Formation Assay for CBD
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Cells (1 × 103) were seeded in each well of a 6-well plate suspended in complete growth media for approximately 16 h, after which time serum free media was used for 6 days for treatment with vehicle control (PBS), CBD, or cisplatin. Colonies were then stained with Giemsa stain containing alcohol for fixation and counted manually by using Leica brightfield microscope (Wetzlar, Germany).
4
Cell Migration Assay with Adipocyte Secretome
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Migration assays were performed using Transwell 8-mm cell culture inserts (BD Falcon, 353097). Briefly, 30,000 to 40,000 cells/well were plated in serum-free medium (SFM) on Transwell filter, and allowed to migrate to medium containing 10% FBS (control media) or AdipoCM. After 8 hours, cells from above the membrane were wiped with cotton swabs, and cells at the bottom were fixed in 10% formalin and stained with 0.05% crystal violet. Cell migration was analyzed by counting cells using a bright field microscope (Leica) and ImageJ. F13A (0.5-4 ng/mL) and etomoxir (40 mmol/L) was added to the cell culture insert at the time of plating.
5
Histological Analysis of Breast Tumor Metastases
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Breast tumor tissues from mice were placed on the sample cassettes and fixed in 10% neutral buffered Formalin (24-72 hours) and transferred into PBS. Samples were embedded in paraffin and 5µm thick sections were taken. Subsequently, the tissue sections were stained using Hematoxylin and Eosin (H &E) and examined for metastases. After incubating at 60° C for one hour, the sections were deparaffinized in Xylene followed by rehydration in graded isopropanol solutions. The slides were then rinsed in tap water and kept in acid alcohol for 3 seconds again followed by tap water wash. Subsequently, the slides were dipped in bluing solution three-four times. Afterwards, the slides were rinsed thrice with tap water and transferred into 70% isopropanol for 3 minutes. Following this, the sections were stained with Eosin Y for 10 minutes. After staining, dehydration was performed in graded isopropanol followed by clearing in Xylene. The slides were then mounted with DPX and were visualized using bright field microscope (Leica).
6
Retinal Vascular Degeneration Measurement
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Ten days after I/R exposure, the remaining rats (5 from each group) were sacrificed to measure degenerate capillaries and pericyte dropout. Briefly, the experimental and control globes are placed into 4% paraformaldehyde for 10 days [15 (link)]. Retina were dissected out and rinsed in water overnight. The retina was then transferred into an elastase solution (40ul/ml) and incubated at 37°C for 2.5h, followed by activation of the elastase in Tris-HCl buffer (pH 8.5) for an additional 12 hours [15 (link)]. The glia and the neuronal regions were gently brushed away. Once only the retinal vasculature was present on the slide, the flatmount was stained with periodic-acid shiff to allow for measurements of degenerate capillaries and pericyte ghosts [1 (link), 15 (link), 16 (link)]. Regions from four retinal flatmounts were counted in each group of animals for pericyte ghosts and degenerate capillaries at ~300μm from optic nerve. Examination was done on 1000 capillary cells (endothelial cells and pericyte) in those 4 fields to count pericyte “ghosts”, excluding any pericyte ghosts observed on degenerate capillaries. Counts were done at 40x using a Leica Brightfield Microscope.
7
Immunohistochemical Analysis of CD31
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Tissues were dissected out; fixed in Bouin’s fixative overnight, cryo-protected and serial sections were cut on a cryostat (CM1850; Leica) at 4–5 μm thickness. The tissue sections were washed with PBS, antigen retrieval was done with enzymatic retrieval method using trypsin-CaCl2 solution. Sections were blocked with 1% BSA at 37 °C. Sections were incubated overnight at 25 °C in a humid atmosphere with primary antibodies against CD31 (1:100; Santa Cruz Biotechnology, Inc.). The sections were and then incubated with biotinylated anti-mouse IgG followed by peroxidase conjugate extra-avidin (Sigma-Aldrich; 1:100) for 60 minutes and 3,3′-diaminobenzidine was used as chromogen (Sigma-Aldrich AEC101-1KT; 1:100) to visualize the reaction product and counterstained with hematoxylin (1:1; Himedia, India). Finally, sections were washed in distilled water and mounted in glycerol gelatin. Kidney and liver sections were cut on a cryostat at 4–5 μm thickness. Sections were stained with haematoxylin and eosin. Images were acquired with a bright-field microscope (Leica) at 10x magnification.
8
Culturing and Visualizing Cancer Stem Cells
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The growth of the cells was monitored on days 1, 3, 6, 9, and 12. On day 12, the spheres were detached from the surface of the flask and floated in suspension as clumps/aggregates. The CSCs bodies thus collected, were visualized under a bright field microscope (Leica, DMIL, Germany), as described by Gu et al.19 (link) Parent HeLa cells (1 × 104 cells per well), included as a control, were cultured in complete DMEM at 37 °C in a 5% CO2 incubator.
9
E-cadherin Immunohistochemistry in Tissue Sections
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Tissues were dissected out; fixed in Bouin’s fixative overnight; cryoprotected in 10% (2 hours), 20% (2 hours), and 30% (overnight) sucrose solution in PBS at 4°C; and frozen with expanding CO2, and serial sections were cut on a cryostat (CM1850; Leica) at 15-μm thickness. The tissue sections were washed in PBS (pH 7.45) for 15 minutes and treated with 1% BSA in PBS containing 0.1% Triton X-100. Sections were incubated overnight at 25°C in a humid atmosphere with primary antibodies against E-cadherin (1:100; Santa Cruz Biotechnology, Inc.) diluted in PBS containing and 1% BSA. Sections were rinsed in PBS for 10 minutes and incubated with biotinylated anti-mouse IgG (Sigma-Aldrich; 1:100) for 1 hour, followed by ExtrAvidin-peroxidase conjugate (Sigma-Aldrich; 1:100) for 40 minutes. 3-Amino-9-ethyl carbazole was used as chromogen (Sigma-Aldrich; 1:100) to visualize the reaction product. Thereafter, sections were counterstained with hematoxylin (1:1; Himedia, Mumbai, India). Finally, sections were washed in distilled water and mounted in glycerol gelatin. Images were acquired with a brightfield microscope (Leica) at 10× magnification.
10
Cardiac Fibrosis Analysis Protocol
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After echocardiographic evaluation, freshly isolated hearts were fixed with paraformaldehyde (4%), embedded in paraffin, and transversally sectioned (thickness, 4–5 μm). For immunohistochemistry (IHC) staining, tissues were incubated with anti-periostin (Abcam, 1 : 200), and the staining was performed as per the methods described previously [18 (link)]. Masson's trichrome and Sirius red staining (Sigma) were performed following the manufacturer's instructions. Staining cross-section heart tissue images were recorded using a bright-field microscope (Leica). To evaluate the degree of cardiac fibrosis, the National Institutes of Health (NIH) ImageJ software was used to analyze images by comparing blue- or red-stained area (collagen) with the total area (10 sections were randomly chosen per heart, n = 6 hearts per group).
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