The scratch assay was used to monitor tumor cell migration. Briefly, 2.5 × 104 MCF-10A neoT cells/well and 8 × 105 MMTV-PyMT cells/well were seeded into a 6-well plate and allowed to adhere and form a confluent layer overnight at 37 °C. The medium was then replaced with fresh SFM or medium, respectively, containing Z9 , nitroxoline, both nitroxoline and Z9 (all 5 µM), or DMSO (0.1%). After 2 h of incubation at 37 °C, the cell monolayer was scraped using a 200 µL pipette tip. Images were acquired at 0 and 16 h (for MCF-10A neoT cells) or 24 h (for MMTV-PyMT cells) on an Olympus CKX 41 light microscope with an Olympus E-450 camera (Olympus, Tokyo, Japan) at 10× magnification. The scratch area was determined using ImageJ software with the wound-healing size tool (https://github.com/AlejandraArnedo/Wound-healing-size-tool/wiki ) [30 (link)]. Cell migration was expressed as a percentage of scratch area and calculated using the following equation: Area (%) = 100 × (At/A0), where At and A0 denote gap area at time t (16 or 24 h) and the initial time point, respectively. For each individual treatment condition, at least two images with different fields of view were acquired and five independent replications of experiment were performed.
E 450 camera
Manufactured by Olympus
Sourced in Japan
The Olympus E-450 is a digital single-lens reflex (DSLR) camera. It features a 10-megapixel sensor and supports interchangeable lenses. The camera is capable of capturing still images and video.
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Lab products found in correlation
4 protocols using e 450 camera
1
Assay for Tumor Cell Migration
2
Wood Degradation Monitoring Methodology
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In order to monitor wood degradation, a GSL-1 sliding microtome (WSL, Switzerland) was used to cut thin wood sections (10-15 μm) of the blocks (20 l ×10 r ×8 t mm 3 ). The sections were stained with safranin (0.5 % aqueous), Astra Blue (0.3 % aqueous) solution and mixed in a 1:1 ratio, washed in distilled water for 1-3 min and dehydrated by an alcohol series. After rinsing in xylol for 1-2 min, sections were mounted in Mountalan glue (Kimianovin, Tehran, Iran) on microscope slides. To avoid buckling of the sample, a 50 g weight was placed on the cover glass edges while the slide was drying at 60 °C for 12 h. Dried sections were examined and photographed with an Olympus E-210 microscope and with an Olympus E-450 camera.
3
Lipid Droplet Imaging in Hepatocytes
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Lipid droplets containing triacylglycerols were observed on hepatocytes cultured for 72h with and without Scia, in DMEM supplemented with 7% SVF and containing 1µmol/L of both insulin and dexamethasone. Lipids were stained with oil red O (CliniSciences, Nanterre, France) following a procedure described by the manufacturer. Cells were observed at a magnification of 40x by an Olympus E-450 camera coupled to an Olympus IX51 microscope (Olympus, Paris, France).
4
Assessing Anti-Migratory Potency of Vaa-Dis
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The wound-healing assay (WHA) was used to estimate the potency of Vaa-Dis to inhibit cell migration (anti-migratory potency). MDA-MB-231 cells were trypsinized, counted as specified above and plated in 48-well cell culture dishes (TPP, Switzerland) at a density of 2 × 10 5 cells/well. Wounds (millimetre gaps) were then scratched in each cell monolayer using a pipette tip. Dead cells were removed by washing with D-PBS buffer and solutions of Vaa-Dis in RPMI-1640/10% FBS at different concentrations (2.5, 5.0, 7.5 and 15.0 nM) were added. Control samples contained only media. The influence of Vaa-Dis on cell migration was determined by observing the width of the gap under a CKX41 inverted microscope equipped with an E-450 camera (Olympus, Japan) after 1 h, 6 h, 12 h and 24 h of incubation. The gap was analysed using the ImageJ software (Softonic International S.A., Spain). The width of the gap was measured as specified (Suppl. Fig. 1) in three different wells for each concentration and incubation time.
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