A wound scratch assay was used to measure the migration of VSMCs. Cells were plated into 6-well culture dishes at a density of 2.0 × 105 cells/well. After adhering, cells were grown to about 80% confluency, then a sterile plastic 10 µL micropipette tip was used to create a 1 mm scratch as a homogeneous wound. Cells were washed twice with phosphate-buffered saline after wounding, and were further incubated for 24 hours. The wound widths were then measured under microscope and images of the wound were then taken using the IX51 inverted microscope (Olympus Optical Co., Tokyo, Japan) at magnification ×100; the wound width was used to calculate cell migration as a wound healing percentage as follows: (0 hours wound width – 24 hours wound width)/0 hours wound width × 100%.
Ix51 inverted microscope
Manufactured by Olympus
Sourced in Japan, United States
The IX51 inverted microscope is a versatile laboratory equipment designed for a range of microscopy applications. It features a stable, ergonomic design and advanced optics to provide clear, high-quality images. The IX51 is a reliable tool for researchers and technicians in various scientific fields.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (10 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (8 mentions)
Xm10 digital camera system, by Olympus (4 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (4 mentions)
Microtome, by Leica (3 mentions)
Hoechst 33342, by Merck Group (3 mentions)
Matrigel, by Corning (3 mentions)
Growth factor reduced matrigel, by BD (3 mentions)
Fluoro gel 2 containing dapi, by Electron Microscopy Sciences (2 mentions)
Dp70 digital camera, by Olympus (2 mentions)
Calcein am, by Thermo Fisher Scientific (2 mentions)
Oil red o, by Merck Group (2 mentions)
Triton x 100, by Solarbio (2 mentions)
Cellsens software, by Olympus (2 mentions)
Dp70 digital camera system, by Olympus (2 mentions)
24 well boyden chamber, by Corning (2 mentions)
Transwell chamber, by Corning (2 mentions)
24 well transwell inserts with 8 μm pore membrane filters, by Corning (2 mentions)
Alamarblue assay, by Thermo Fisher Scientific (2 mentions)
Live dead assay, by Thermo Fisher Scientific (2 mentions)
178 protocols using ix51 inverted microscope
1
Wound Scratch Assay for VSMC Migration
2
Quantifying Microcystis Growth Dynamics
3
Immunofluorescence Staining Protocol
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Cells were fixed in 4% paraformaldehyde for 30 min, permeabilized in 0.1% Triton X-100 for 10 min, and blocked with 3% bovine serum albumin (BSA) for 30 min at room temperature. The cells were then incubated with the appropriate antibodies overnight at 4°C, followed by incubation for 1 h at room temperature with secondary fluorescent antibodies. Finally, the cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen) for 5 min to label the nuclei. Images were acquired using an Olympus IX51 Inverted Microscope (Olympus Optical)
4
Cell Migration Assay Protocol
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SH-SY5Y cells were seeded into 24-well cell culture plates and were allowed to grow to 100% confluence as a monolayer. The monolayer was gently scratched across the center of the well with a sterile pipette tip. After scratching, the medium was removed, and the wells were washed twice in PBS solution. Fresh medium containing no FBS and designated treatments were added to each well. Images were obtained from the same fields immediately after scratching (t0) and 48 h later. The scratch was visualized by phase-contrast light microscopy using an Olympus IX51 inverted microscope (Olympus Optical Co., Hamburg, Germany). The images were acquired in randomly selected fields by using an Olympus digital camera and analyzed by ImageJ software (National Institutes of Health, Bethesda, MD, USA). The percentage of the closure of the scratch was then calculated.
5
Intestinal Morphology Measurement Protocol
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Samples were dehydrated, cleared, and embedded in paraffin. Intestinal segments from 6 hens per treatment were sectioned to obtain samples of 5-μm thickness, which were placed on glass slides and stained with hematoxylin and eosin as per standard paraffin-embedding procedures for examination by light microscopy (Forte et al., 2016 (link)). Villus height was measured from the tip of the villus to the villus-crypt junction, and crypt depth was determined from the base up to the crypt-villus transition region. Morphological measurements were performed on 10 villi chosen from each segment, using an image processing and analyzing system (version 6.0, Olympus IX51 inverted microscope, Olympus Optical Co., Ltd., Tokyo, Japan). The villus height-to-crypt depth ratio was subsequently calculated and recorded.
6
Intestinal Histomorphometry: Villus and Crypt Analysis
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Formalin-fixed intestinal tissues were processed, dehydrated, embedded in paraffin wax, sectioned at 3 μm and stained, using the hematoxylin and eosin method. Histological sections were examined with Villus height (VH ), villus width (VW ), and crypt depth (CD ). Morphological measurements were performed on 10 villi chosen from each segment, using an image processing and analyzing system (version 6.0, Olympus IX51 inverted microscope, Olympus Optical Co., Ltd., Tokyo, Japan). The villus height-to-crypt depth ratio (VH/CD ) was calculated subsequently.
7
Intestinal Histomorphometry Analysis
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Intestinal tissue samples were dehydrated, cleared, and embedded in paraffin. Serial sections were then cut at 5um thickness, deparaffinized in xylene, rehydrated, and stained with hematoxylin and eosin as per standard paraffin embedding procedures for examination using a microscope. Villus height and crypt depth of 10 welloriented villi per segment were measured using an image processing and analyzing system (Olympus IX51 inverted microscope, Olympus Optical Co., Ltd., Tokyo, Japan). The villus height-to-crypt depth ratio was subsequently calculated.
8
Ovarian Cancer Tissue Microarray Analysis
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Human ovary tissue microarrays (OV1005a and OV808) were purchased from the US Biomax, Inc. (Alenabio Biotechnology, Xi'an, China). The microarrays contained tissues from 3 normal ovaries, 17 cancer adjacent normal ovaries, 18 benign cystadenomas, 7 borderline cystadenomas, 45 primary malignant epithelial ovarian cancers, as well as 50 metastatic ovarian cancers. Immunohistochemistry for ROR2 was performed as described previously. 23 All the samples were observed and photographed with the Olympus IX51 Inverted Microscope (Olympus Optical, Melville, NY, USA). The product of the intensity scores was used as the final ROR2 staining score. Staining intensity was scored as 0, 1, 2, or 3 (no staining, weak, moderate, or strongly positive). The final evaluation criteria of the ROR2 expression was designated as negative or positive (negative: 0-1, positive: 2-3). All the scores of ROR2 expression were determined independently by two senior pathologists.
9
Live/Dead Assay for Microtissues
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A live/dead assay is performed using a combination of standard viability stains, calcein AM (acetoxymethyl) and propidium iodide (PI, both Invitrogen). First, live microtissues are stained using calcein AM. Before staining, the medium is carefully drained from the Petri dish and tissue samples are washed twice with PBS. Culture medium containing 1 μg/mL of the calcein AM is added to the Petri dish. After 1 h incubation at 37°C, the medium is removed and the sample washed twice with PBS. To induce cell death, microtissues are exposed to a 50% ethanol solution in culture medium for 45 min. After washing twice with PBS, culture medium supplemented with 1 μg/mL PI is added to the Petri dish. After 1 h incubation at 37°C and washing with PBS twice, the sample is imaged using fluorescence microscopy (IX51 Olympus inverted microscope equipped with a X-cite 120 PC Hg lamp using a 2x objective).
10
Inflammasome Activation in BMMs
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BMM were plated at 5 × 104 cells per well on a 96-well plate and maintained for 24 h in culture media. They were primed with 100 ng/ml LPS for 3 h and treated with 10 µM CDD-450 and/or 15 µM nigericin for 30 min. Cells were incubated with the FLICA FAM-YVAD-FMK probe (ImmunoChemistry Technologies) for 30 min at 37°C, washed twice with PBS/FBS, and fixed with 10% buffered formalin. Cells were then counterstained with Fluoro gel II containing DAPI (Electron Microscopy Sciences). Images were taken with a DP70 Olympus digital camera coupled with an IX51 Olympus inverted microscope, captured with QCapture Pro software (QImaging), and analyzed with ImageJ software (National Institutes of Health).
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