Ckx52 microscope

Manufactured by Olympus

The CKX52 is an inverted microscope designed for routine microscopy applications. It features a compact and ergonomic design, a stable and rigid frame, and a variety of optical components to support basic observation and imaging tasks.

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Lab products found in correlation

Matrigel coated filter, by Corning (1 mentions) Cytation 5 cell imaging multi mode reader, by Agilent Technologies (1 mentions) Gemcitabine, by Selleck Chemicals (1 mentions) Human recombinant epidermal growth factor, by Thermo Fisher Scientific (1 mentions) Ultra low cluster plate, by Corning (1 mentions) Recombinant human basic fibroblast growth factor, by Thermo Fisher Scientific (1 mentions) G fectin, by Genolution (1 mentions)

4 protocols using ckx52 microscope

Transwell Assay for Cell Migration and Invasion

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A transwell assay was performed to estimate migration and invasion abilities. Briefly, DU145, PC-3, and LNCap cells transfected with the control miRNA or miR-31-3p were suspended in serum-free RPMI-1640 medium. The suspended cells were reseeded at a density of 10⁵ cells/well onto filters or Matrigel-coated filters in the upper chamber of an 8.0-μm-pore transwell plate (Corning Inc.). After incubation for 72–96 h, the migrated and invaded cells on the lower surface of the filters were fixed in methanol and stained using 0.05% crystal violet solution. The stained cells were counted in four randomly selected fields using an Olympus CKX52 microscope (magnification, ×200). Experiments were performed in triplicate and repeated three times.

Choi S., Lee S., Han Y.H., Choi J., Kim I., Lee J, & An H.J. (2022). miR-31-3p functions as a tumor suppressor by directly targeting GABBR2 in prostate cancer. Frontiers in Oncology, 12, 945057.

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Vascularized Organoid Drug Response

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Vascularized organoid and co-culture tissues were created on InVADE platform as described above. Eight days post-seeding, 1 μM gemcitabine (SelleckChem) in endothelial growth medium was added to the perfusion media in 3D culture. Four days later (12 d), cell growth was analyzed using CellTiter-Glo 3D as described above.
As static controls, same cell density primary pancreatic organoid and organoid-fibroblast co-culture (~14,000 total cells on day 0) were seeded as 3D Matrigel domes on a 96 well plate, and similarly treated eight days post-seeding with 0, 0.01, 0.1 or 1μM gemcitabine diluted in OGM medium. The cell proliferation of the tissues maintained under static conditions was also assessed using CellTiter-Glo 3D cell proliferation assay (12 d) and the luminescence was captured with Cytation5 Cell Imaging Multi-Mode Reader (Biotek).
Bright-field images of the 4-day drug treatment on both perfused inVADE tissues and static 96-well plate tissues were captured with the Olympus CKX52 microscope (4X objective).

Lai Benjamin F.L., Lu Rick X., Hu Y., Davenport H.L., Dou W., Wang E.Y., Radulovich N., Tsao M.S., Sun Y, & Radisic M. (2020). Recapitulating pancreatic tumor microenvironment through synergistic use of patient organoids and organ-on-a-chip vasculature. Advanced functional materials, 30(48), 2000545.

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Sphere Formation Culture and Transfection

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For sphere formation culture, the cells were incubated in Dulbecco’s modified eagle’s/F12 serum-free medium (Welgene Inc.) supplemented with 2% B-27 (Gibco; Thermo Fisher Scientific Inc.), 20 ng/mL recombinant human epidermal growth factor (Gibco; Thermo Fisher Scientific Inc.), and 20 ng/mL recombinant human fibroblast growth factor basic (Gibco; Thermo Fisher Scientific Inc.) in six-well ultra-low cluster plates (Corning Inc.). Once the diameters of the formed spheres reached 50 µm, the spheroids were transfected with miR-31-3p or control miRNA at a final concentration of 80 nM using G-fectin (Genolution Pharmaceuticals Inc.) according to the manufacturer’s protocol. Four days after transfection, images of the spheres were captured using an Olympus CKX52 microscope (magnification, ×40), and the number and diameter of the spheres were measured in four randomly selected fields. Experiments were performed in triplicate and repeated three times.

Choi S., Lee S., Han Y.H., Choi J., Kim I., Lee J, & An H.J. (2022). miR-31-3p functions as a tumor suppressor by directly targeting GABBR2 in prostate cancer. Frontiers in Oncology, 12, 945057.

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Transwell Assay for Cell Migration and Invasion

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Cells were suspended in a serum-free RPMI-1640 medium. The suspended cells were seeded at a density of 10⁵ cells/well to the filter (with or without Matrigel) in the upper chamber of 8.0-μm transwell plate (Corning). After 72-96 h, migrating and invading cells on the lower surface of the filter were fixed in methanol and stained with 0.05% crystal violet solution. The stained cells were counted in four randomly selected fields of view using Olympus CKX52 microscope (magnification, × 200).

Luo L., Li P., Xie Q., Wu Y., Qin F., Liao D., Zeng K, & Wang K. (2024). n6-methyladenosine-modified circular RNA family with sequence similarity 126, member A affects cholesterol synthesis and malignant progression of prostate cancer cells by targeting microRNA-505-3p to mediate calnexin. Journal of Cancer, 15(4), 966-980.

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