Ten microliters of sperm fraction was placed on a pre-warmed slide with pre-warmed cover slip and examined quickly for sperm motility. The slides were visualized at 200× with a temperature-controlled (37°C) phase-contrast BX 60 Olympus microscope equipped with a digital imaging camera (DP-12) and Micro Image Lite software 4.0 (Olympus). At least four fields were evaluated per group. Spermatozoa from caput and cauda epididymides were counted using Neubauer's hemocytometer. For sperm morphology, spermatozoa were collected immediately after killing of mice by flushing the vas deferens with phosphate-buffered saline (pH 7.2), and a drop was smeared on a pre-warmed slide with pre-warmed cover slip. The slides were visualized at 1000× with a temperaturecontrolled (37°C) phase-contrast BX 60 Olympus microscope as described earlier.
Bx60 microscope
Manufactured by Olympus
Sourced in Japan, United States, Germany, United Kingdom, Canada, France, Australia, Sweden
The BX60 microscope is a high-performance optical microscope designed for a variety of laboratory applications. It features a sturdy, ergonomic design and advanced optical components that provide clear, high-quality images. The BX60 is capable of both transmitted and reflected light observation, making it suitable for a wide range of specimen types.
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Lab products found in correlation
Dp71 camera, by Olympus (12 mentions)
Dp72 camera, by Olympus (12 mentions)
Dp70 digital camera, by Olympus (10 mentions)
Dp70 camera, by Olympus (8 mentions)
Lsm 700 confocal microscope, by Zeiss (8 mentions)
Dp71 digital camera, by Olympus (7 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (6 mentions)
Cell f software, by Olympus (6 mentions)
Image pro plus 6, by Media Cybernetics (6 mentions)
Cresyl violet, by Merck Group (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (5 mentions)
Ultracut uct ultramicrotome, by Leica (4 mentions)
Dp70 camera unit, by Olympus (4 mentions)
Paraformaldehyde (pfa), by Merck Group (4 mentions)
Mitotracker red cmxros, by Thermo Fisher Scientific (4 mentions)
Cellsens software, by Olympus (4 mentions)
In situ cell death detection kit, by Roche (3 mentions)
Signalstain boost ihc detection reagent, by Cell Signaling Technology (3 mentions)
Fv1000 confocal microscope, by Olympus (3 mentions)
Lsm 780, by Zeiss (3 mentions)
531 protocols using bx60 microscope
1
Sperm Motility and Morphology Analysis
2
Immunofluorescence Staining of Murine Eye Tissues
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Mice were euthanized, and eyes collected and fixed in 2% paraformaldehyde for 1 hr before transferring into cold PBS. For staining of endothelial or pericyte markers eyes were fixed in ice cold methanol. After dissection of the various compartments of the eye (choroid, retina, iris) the tissues were incubated in 20 mM EDTA at 37°C for 30 minutes. Tissues were then incubated in a solution containing 0.3% Triton-X, 2% bovine serum albumin and 10% of normal goat serum in PBS at room temperature for 30 min. Tissues were incubated with the primary antibody overnight at 4°C followed by incubation with the secondary antibody at room temperature for 1 hr. Detection of the immediate-early (IE1) protein of MCMV was performed with the 6/58/1 monoclonal antibody [53 (link)]. Monoclonal antibodies specific for the following cell surface markers were used to stain tissue sections: MHC-class II (clone M5/114), CD45 (clone 30F11), CD31 (clone Mec 13.3), PDGFRβ (clone APB5), F4/80 (clone BM8), CD4 (clone RM4-5), CD8 (clone 53–6.7), IBA1 (Wako Pure Chemicals Industry, Osaka, Japan), CollagenIV (Biorad-2150-1470). Slides were counterstained with Hoechst to visualize nuclei. Assessment of stained specimens was performed using an epifluorescence microscope (Olympus BX60 microscope: Olympus, Tokyo, Japan) or a Nikon C2 Upright Confocal microscope.
3
Immunofluorescence Staining of T. cruzi
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T. cruzi expressing MSH6 fused to mRFP or to 6x HA epitopes were fixed with 4% paraformaldehyde for 5 min, permeabilized with 0.1% Triton X-100 for 10 min, blocked with 1% BSA, 0.2% Tween 20 for 1 h at room temperature and incubated with 1:500 anti-HA antiserum (Roche) for 1 h. After washing with PBS, nuclei were stained with 1 μg/mL of DAPI (Molecular Probes/ Life Technologies) for 5 min and cover slides mounted with prolong gold anti-fade solution (Molecular Probes/Life Technologies). Images were acquired with a 100x objective in the fluorescence microscope Olympus BX60 microscope using Q-color 5 digital camera and Qcapture Pro 6.0 software and with Nikon Eclipse Ti Tecnai G2-12 SpiritBiotwin FEI (120kV) at the Centro de Aquisição e Processamento de Imagens (CAPI-ICB/UFMG).
4
Pancreatic beta cell quantification
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Rats’ pancreases were fixed in aqueous Bouin solution for 24 h and embedded in paraffin (Labonord, Templemars, France). The entire pancreas was homogeneously sampled using a fixed-interval sampling method to avoid bias due to regional change in islets distribution, as previously described [24 (link)]. Briefly, each paraffin block was serially sectioned (5 μm) throughout its full-length, sections at a fixed interval were mounted on slides and stored at room temperature for future immune-staining.
Pancreatic sections were blocked for 1 h with 10% normal goat serum in Tris buffer saline and then probed for insulin using primary mouse anti-insulin antibody (Santa Cruz, France) (1:200). Sections were then incubated with an HRP conjugated anti-mouse secondary antibody (Jackson immunoResearch Laboratories, Ely, UK) (1:200). β cells areas were determined by morphometric analysis in pancreatic sections stained for insulin, using an OLYMPUS BX60 microscope equipped with Histolab 10.5.1 computer-assisted image analysis system software (Microvision Instrument, Evry, France). The relative β cell area in each stained section was calculated as the ratio of insulin positive cells area over the total pancreatic cells area [24 (link),25 (link)]. At least six sections homogeneously distributed were analyzed for each pancreas.
Pancreatic sections were blocked for 1 h with 10% normal goat serum in Tris buffer saline and then probed for insulin using primary mouse anti-insulin antibody (Santa Cruz, France) (1:200). Sections were then incubated with an HRP conjugated anti-mouse secondary antibody (Jackson immunoResearch Laboratories, Ely, UK) (1:200). β cells areas were determined by morphometric analysis in pancreatic sections stained for insulin, using an OLYMPUS BX60 microscope equipped with Histolab 10.5.1 computer-assisted image analysis system software (Microvision Instrument, Evry, France). The relative β cell area in each stained section was calculated as the ratio of insulin positive cells area over the total pancreatic cells area [24 (link),25 (link)]. At least six sections homogeneously distributed were analyzed for each pancreas.
5
Seminal Vesicle Contamination Analysis
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To observe contaminated and uncontaminated seminal vesicles in whole mount, adult males were dissected and seminal vesicles removed in 0.1 M sodium phosphate buffer solution, pH 7.2, fixed for 1 h in 4% paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.2 and transferred to histological slides covered with coverslips and photographed using an Olympus BX-60 microscope equipped with phase contrast (Olympus Corporation, Tokyo, Japan).
Some isolated seminal vesicles were dissected out and the parasites and spermatozoa were spread on slides, fixed with solution of 4% paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.2 for 30 minutes, washed in running water and dried at room temperature. The slides were examined and the parasites were photographed in Olympus BX-60 photomicroscope equipped with phase contrast. To observe the nuclei some slides were stained with 0.2 mg/ml4,6-diamino-2-phenylindole (DAPI).
Some isolated seminal vesicles were dissected out and the parasites and spermatozoa were spread on slides, fixed with solution of 4% paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.2 for 30 minutes, washed in running water and dried at room temperature. The slides were examined and the parasites were photographed in Olympus BX-60 photomicroscope equipped with phase contrast. To observe the nuclei some slides were stained with 0.2 mg/ml4,6-diamino-2-phenylindole (DAPI).
6
Golgi-Cox Staining of Nicotine Effects
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Nicotine-treated rats and vehicle controls were deeply anesthetized with Allfatal (100 mg/ml (pentobarbital); Omnidea AB, Stockholm, Sweden) and perfused intracardiovascular with Tyrode's solution and 4% para-formaldehyde. Brains were impregnated in a Golgi-Cox Fast Kit according to the company protocol (FD NeuroTechnologies, Columbia, MD), and cut into 60 μm slices. A Zeiss lsm 700 inverted confocal microscope with a × 63 oil objective (NA 1.4), and an Olympus BX60 microscope (Olympus Corp., Center Valley, PA) equipped with a × 100 oil objective (NA 1.3) were used to capture images from the distal dendrites. Spine counting was carried out blind to treatment on at least two separate occasions/treatment.
7
Immunohistochemical Analysis of Tumor Infiltrates
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The grafted tumor masses were fixed in 10% formalin, embedded in paraffin and sectioned with a thickness of 3 µm for immunohistochemical studies. Rabbit anti-human CD3 polyclonal antibody (cat. no. ab828; dilution, 1:50; Abcam, Cambridge, MA, USA) and mouse anti-human perforin monoclonal antibody (cat. no. ab47225; dilution, 1:50; Abcam) were used for immunostaining. Horseradish peroxidase-conjugated goat anti-rabbit (cat. no. ab6721; dilution, 1:1,000; Abcam) and goat anti-mouse (cat. no. ab6789; dilution, 1:1,000; Abcam) secondary antibodies were used for detection of the primary antibodies. All procedures were conducted according to the manufacturer's protocol. Images of the sections were acquired using an Olympus BX-60 microscope (Olympus Corporation, Tokyo, Japan) to determine the CD3+ and perforin+ cell densities. The number of positive cells was blindly counted in 10 randomly selected independent fields (0.16 mm2 at ×400 magnification) for each section by two independent observers.
8
Corneal Histology of Klf5-Deficient Mice
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Klf5Δ/ΔCE and control mice were euthanized via carbon dioxide asphyxiation and cervical dislocation, and their eyes were immediately fixed in 4% paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in PBS (pH 7.4) for 24 hours at room temperature (RT). Whole globes were embedded in paraffin, and central corneal 5-μm sections were stained with hematoxylin and eosin (H&E) or periodic acid Schiff (PAS) reagent following standard protocols. Sections were viewed with an Olympus BX60 microscope (Olympus America, Inc., Allentown, PA, USA) and captured using a Spot digital camera (Spot Diagnostics Instruments, Inc., Sterling Heights, CA, USA). All images were processed similarly using Adobe Photoshop and Illustrator (Adobe Systems, San Jose, CA, USA).
9
Evaluating MPC Migration on Fibrin Filler
10
Apoptosis Detection in Tumor Sections
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Tumor samples were fixed with 4% paraformaldehyde for 4 h, and then dehydrated by graded sucrose solution. Subsequently, the tumor samples were embedded in Tissue-Tek Optimal Cutting Temperature compound (Sakura, Alphen aan den Rijn, The Netherlands), and 10-µm frozen sections were cut sagittally with a freezing microtome. Apoptosis was detected by the In Situ Cell Death Detection kit (Roche Applied Science, IN, USA). Briefly, frozen sections were incubated with permeabilization solution for 2 min on ice. After washing, the sections were incubated with the TUNEL reaction mixture for 60 min at 37°C in the dark and then rinsed with PBS. Subsequently, the sections were sealed with VECTASHIELD mounting medium with DAPI (Vector Laboratories, Inc., Burlingame, CA, USA) and visualized under the Olympus BX60 microscope (Olympus Corporation, Shinjuku, Japan).
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