Apricell 3-in-1 Spheroid Organoid Plate
Each insert has 24 spheroids (4 different quadrants with 6 spheroids/organoids per quadrant). Each quadrant is separated from one another, so you can run 4 different experimental conditions in the one insert.
There will only ever be one spheroid or organoid per microwell.
The hydrogel used to form the 3-in-1 Plate is non-cell adherent. Occasionally cells will become stuck in the channels that lead to the microwells. If this occurs, gently rinse any additional cells stuck in the channels by adding 250-300 μL of pre-warmed medium to the Cell & ECM Loading Zone. In most instances, cells that become stuck in the channels will naturally slide down the channels during the incubation period over the course of a few hours.
There may be a small amount of media left in the microwells after pipetting from the Media Reservoir. If this occurs, simply aspirate the remaining media from the Cell & ECM Loading Zone.
Yes. The plate can be placed on ice. We do not recommend placing it on ice for more than 20 minutes at a time. Freezing our platform is not recommended as it can distort the delicate microwell structure and reduce spheroid consistency.
The size of the spheroids/organoids is determined by a variety of factors: the quantity of cells that are seeded, the type of cells, the size of the microwells and the duration of culture.
Adding an ECM hydrogel is not always necessary, depending on the type of model you wish to create. Spheroids can be formed without a hydrogel. However, culturing primary cells to form organoids typically requires an ECM hydrogel.
The Secondary Reservoir has been optimized for certain co-culture and tri-culture experiments. For example, fibroblast co-culture with tumor cells. It is also ideal for experiments that require a secondary ECM, such as in many tri-culture experiments. For these types of experiments, a primary ECM should be added to the Cell & ECM Loading Zone, and a secondary ECM overlay should be added to the Secondary Reservoir.
The 3-in-1 Plate is compatible with all standard forms of microscopy: inverted, upright, bright field, fluorescent, confocal, etc.
Yes. In fact, an inverted microscope is ideal. For improved imaging the inserts can be removed from the well and placed on a coverslip.
No. Users have not reported any distortion/reflection for imaging.
Yes, as long as it doesn’t affect cell proliferation.
Yes. The 3-in-1 Plate is compatible with confocal microscopy and all staining can be done within the insert. For getting higher resolution images in confocal imaging, we recommend removing the insert from its well using a lab spatula and placing it on the coverslip. To remove the insert, press the lab spatula into the gap between the insert and the plastic of the well plate. Once at the bottom of the six-well plate, flex your lab spoon towards the center, this motion should pop the insert out of the well. Securely support the insert with the lab spatula and remove from the six-well plate.
The hydrogel type, cross-linking and concentration and time are all factors that can impact the matrix stiffness. What ECM hydrogel you choose for your experiment is ultimately up to you. The 3-in-1 Plate should be compatible with whatever you end up choosing. Ensure that the viscosity of the hydrogel is not excessively viscous when pipetting into the Cell & ECM Loading Zone.
All staining can be done on the plate. There is no need to remove the spheroids or even to digest the extracellular matrix.
No, the inserts are press fit into the well and are designed not to move even when media is added. The inserts are designed with tight tolerances to the exact dimensions of the well such that friction holds the inserts in place.
The wall design between the quadrants prevents media and added compounds from mixing from one to another quadrant. Cross-contamination may occur if the plate were bumped or too much media added, spilling media over the barrier into other quadrants.
By following the Cell & ECM Seeding Protocol, cells will evenly distribute across the microwells. For diluted cell suspension, for example, at < 50 k cells/50uL), it is recommended to keep the pipette tip close to the microwell section for even cell distribution.
The Apricell inserts provide a ULA material to grow spheroids. The design enables disturbance-free media changes, co-culture ability, and a flat bottom surface which helps generate images with increased clarity when compared to round bottom and hanging drop plates. A total of 144 spheroids may be produced in each plate, with six replicates per isolated quadrant. In addition, without transferring from the spheroids to other plates, extracellular matrices may be added directly to tumoroids and the insert may directly be sliced for immunohistochemistry analysis.
