Get Perfecta3d Hanging Drop Plate Size 96 Wells , With Lid And Tray ...

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2. Begin by reviewing the required materials for the assay, ensuring you have the Perfecta3D Cellular Invasion Assay Kit and healthy proliferating cells optimized for spheroid formation.
3. Fill the peripheral rim reservoirs of the plate and tray with 2 mL per plate reservoir section and 1 mL per tray reservoir section of water or buffer. Optionally, prepare an agarose solution if needed.
4. If exogenous extra cellular matrix (ECM) is required, thaw 10X Spheroid Formation ECM at 4°C and prepare a cell suspension in 1X Spheroid Formation ECM at a concentration of 2000 to 5000 cells.
5. Pipet the cell suspension into each well of the Hanging Drop Plate to form drops of 30 to 50 µL, ensuring they remain on the bottom side of the plate.
6. Cover the assembly with the lid and incubate it until individual cells form into spheroids, typically within 72 hours.
7. Prepare the Invasion Matrix by thawing it at 4°C and adding 50 µL to a pre-chilled 96-well U-bottom invasion assay plate.
8. Transfer the Hanging Drop Plate to the U-bottom plate carefully and centrifuge both plates to eliminate bubbles and settle spheroids.
9. After one hour at 37°C to promote polymerization, introduce 100 µL of warm cell culture medium containing any required compounds.
10. Incubate the invasion assay for 3 to 6 days, photographing the spheroids every 24 hours to analyze morphological changes.

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