For cell proliferation, cells were cultured in 96-well plates and placed in the IncuCyte live-cell imaging system (Essen BioScience, USA). Cells were observed using phase microscopy (NiKon, #MRH00101) and calculated by IncuCyte analysis software. For colony formation, cells were seeded at a density of 500 cells per well in 6-well plates and incubated. Two weeks later, cells were fixed with 4% paraformaldehyde and stained by crystal violet. The images of the colonies were captured, and the numbers of the colonies were counted by the software Image-Pro Plus 6.0.
Mrh00101
Manufactured by Nikon
Sourced in Canada
The MRH00101 is a laboratory equipment product manufactured by Nikon. It serves as a core component for various scientific and research applications. The device's primary function is to provide precise measurement and analysis capabilities within a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Incucyte live cell imaging system, by Sartorius (3 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Copas biosort, by Union Biometrica (1 mentions)
Mrd00025, by Nikon (1 mentions)
Fluorescent polychromatic beads, by Polysciences (1 mentions)
Imagexpress nano, by Molecular Devices (1 mentions)
4 protocols using mrh00101
1
Cell Proliferation and Colony Assays
2
Quantitative Live-Cell Imaging and Colony Assay
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The IncuCyte live-cell imaging system (Essen BioScience) was utilized initially for the purpose of measuring the rate of cell proliferation. Phase microscopy (NiKon, #MRH00101) was used to capture cell proliferation images and curves after the cells had been grown in 96-well plates and put in the IncuCyte system. This was followed by analysis using the IncuCyte analysis software. To complete the MTS assay, the cells were plated at a density of 5,000 cells/well. Following a seeding duration of 1-3 days, the cells were subjected to 40 minutes of incubation with MTS (Promega, Madison, WI) for 40 minutes at each of those time points. Additionally, with the aid of a microplate reader, the optical density (OD) was measured. To facilitate the formation of colonies, the cells were planted in 6-well plates at 500 cells/well. After incubating the cells for two weeks, 4% paraformaldehyde was utilized to fix them before staining them with crystal violet. Images of the colonies were captured and processed with Image-Pro Plus 6.0 (Media Cybernetics, MD, USA) to facilitate the counting of the total colonies.
3
Live-Cell Imaging and Clonal Analysis
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Cells were cultured into 96‐well plates and placed in the IncuCyte live‐cell imaging system (Essen Bio Science). Cells were observed using phase microscopy with 10× objective (NiKon, #MRH00101) and calculated by IncuCyte analysis software. In addition, 50 cells were cultured into 6‐well plates and cultured for 7 days. Then the cell clones were fixed with 4.0% formaldehyde for 15 minutes and stained with 0.1% purple crystal. The results of cell clones were detected by ChemiDoc MP Imaging System (Bio‐Rad).
4
Quantifying Ingested Particles in Nematodes
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Flasks were sampled each hour beginning one hour after feeding and continuing for 72 consecutive hours. At each hour, 500 µL was removed from each flask and transferred to a well of a deep 96-well plate. Each flask was sampled at each time point. Fluorescent polychromatic beads (Polysciences, 19507-5 ) with a 0.5 μm particle size were added to each well at a final concentration of 3.64x10 8 beads/mL and incubated at 20°C for 10 minutes with shaking. Following the bead incubation, 30 µL from each well of the deep 96-well plate was aliquoted to a 96-well microtiter plate. The process was repeated 11 times to 11 separate wells of the same microtiter plate with pipetting to mix the well contents from the deep 96-well plate. Animals were then treated with sodium azide at a final concentration of 50 mM to paralyze and prevent defecation of the ingested beads.
The 96-well plate was imaged with an ImageXpress Nano (Molecular Devices, SanJose, CA) using both 2x (Nikon MRD00025) and 10x (Nikon MRH00101) objectives. The ImageXpress Nano acquires brightfield images using a 4.7 megaPixel CMOS camera. Images are stored in 16-bit TIFF format. Finally, animals were scored using a large-particle flow cytometer (COPAS BIOSORT, Union Biometrica, Holliston MA). The COPAS BIOSORT sheath flow rate was kept at a constant 10.3 ±0.1 mL per minute to reduce variability in length measurements.
The 96-well plate was imaged with an ImageXpress Nano (Molecular Devices, SanJose, CA) using both 2x (Nikon MRD00025) and 10x (Nikon MRH00101) objectives. The ImageXpress Nano acquires brightfield images using a 4.7 megaPixel CMOS camera. Images are stored in 16-bit TIFF format. Finally, animals were scored using a large-particle flow cytometer (COPAS BIOSORT, Union Biometrica, Holliston MA). The COPAS BIOSORT sheath flow rate was kept at a constant 10.3 ±0.1 mL per minute to reduce variability in length measurements.
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