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Order Now | Features | Applications | Data | Cell Types | Protocols | FAQ
Frequently Asked Questions
1. Plate Re-Use | Can the Perfecta3D™ Hanging Drop Plates be re-used?
2. Evaporation | How can I minimize evaporation of the hanging drops?
3. Extracellular Matrix | Can I use extracellular matrix gels with the Perfecta3D™ Hanging Drop Plates?
4. Cell Types | What cell types can I grow into spheroids using the Perfecta3D™ Hanging Drop Plates?
5. Hanging Drop Formation | How much media does each hanging drop hold?
6. Hanging Drop Formation | How can I form hanging drops more consistently with manual pipetting?
7. Reservoirs | How can I minimize spilling of liquids from the reservoirs on the Plate and Tray?
8. Spheroid Formation | Why aren’t my cells forming spheroids?
9. Cell Treatment | How do I treat spheroids with test compounds?
10. Cell Harvest | How do I harvest spheroids from the Perfecta3D™ Hanging Drop Plates?
11. Automation | How do I set up my liquid handling system to perform automated pipetting?
12. Intellectual Property | What is the intellectual property status of your products?
13. Plate Reader | How do I set up my plate reader to acquire measurements directly from a Perfecta3D™ Hanging Drop Plate?
1. Can the Perfecta3D™ Hanging Drop Plates be re-used?
We do NOT recommend re-using the Perfecta3D™ Hanging Drop Plates. Re-using the plates will introduce uncontrolled variables into your experiments which may affect the outcome of your study. The Perfecta3D™ Hanging Drop Plates are designed for single-use experiments.
2. How can I minimize evaporation of the hanging drops?
Evaporation can be significantly reduced by adding water, buffer solution, or agarose solution to the reservoirs on periphery of the Plate and Tray. Alternatively, or concurrently, hanging drops of water, buffer solution, or agarose solution can be formed in the peripheral access holes to reduce evaporation and edge effect. Please also see #7.
Another method is to replace the bottom tray with a BD Falcon culture plate filled with water, buffer solution, or agarose solution. The bottom tray can be used when conducting light microscopy. BD Falcon plates are recommended because they have same notch positions as the Perfecta3D™ Hanging Drop Plates.
3. Can I use extracellular matrix gels with the Perfecta3D™ Hanging Drop Plates?
Yes, extracellular matrix (ECM) gels, such as Matrigel and collagen, can be used with the Perfecta3D™ Hanging Drop Plates. Simply mix your cell solution with your choice of ECM gel and seed the mixture into the plate. ECM gels can be used at low concentrations as supplement to the culture media. ECM gels can also be used at high concentrations (eg. 50/50 mixture with cells) to form hanging drops of solid gels. Maintain the solid hanging drops by adding 2-3 μL per day or 5 μL every other day (adjust as necessary).
4. What cell types can I grow into spheroids using the Perfecta3D™ Hanging Drop Plates?
Our users have successfully demonstrated spheroid formation using many cell types. You can find the full list here. This list is updated frequently as users share their new findings with us.
5. How much media does each hanging drop hold?
Using the manual pipetting method described here, each drop can hold 20 to 30 μL.
6. How can I form hanging drops more consistently with manual pipetting?
For instructions on manual pipetting, please refer to the guide here.
7. How can I minimize spilling of liquids from the reservoirs on the Plate and Tray?
Water or buffer solution can be replaced with agarose gel to minimize spilling. First, prepare agrose solution by adding agarose to water or buffer solution (eg. 0.1 g to 10 mL water). Melt agarose solution using a microwave. Dispense agarose solution to the reservoirs once the solution has cooled to approximately 50 °C. Finally, allow agarose gel to cool and solidify.
8. Why aren’t my cells forming spheroids?
Not all cell types form spheroids readily when cultured in hanging drops. Please check relevant literature to see if spheroid formation with your cell type of interest has been demonstrated before. A list of working cell types submitted by our users can be found here. Media composition is an important factor to spheroid formation, so make sure your culture media contains the necessary supplements. Some cell types form small clusters of cells initially that later aggregate into one larger spheroid after a few days in culture.
9. How do I treat spheroids with test compounds?
Cell treatments can be easily applied from the topside of the Perfecta3D™ Hanging Drop Plate. An example protocol is to prepare your treatment solutions at two times the final concentrations. Remove 15 μL of media from each 30 μL hanging drop, then add 15 μL of test compound-containing media to each drop.
10. How do I harvest spheroids from the Perfecta3D™ Hanging Drop Plates?
Spheroids can be easily harvested from either the top or bottom side of the Perfecta3D™ Hanging Drop Plate. Once harvested, the spheroids can be processed for further culturing (e.g. invasion assay) or analysis (e.g. flow cytometry, immunofluorescence staining, histological staining, etc.).
To harvest spheroids from the topside, first pre-wet pipette tips with culture media to prevent spheroids sticking to tips. Carefully aspirate and transfer the content to a container (e.g. 15 mL tube) or a multi-well plate (e.g. 384- or 96- well plate).
To harvest spheroids from the bottom side, stack the cell-containing Perfecta3D™ Hanging Drop Plate on top of a receiving vessel (e.g. 384- or 96- well plate). Add just enough media or buffer solution (~40 μL) to each hanging drop to force the drop fall into the vessel below.
To selectively harvest spheroids from the bottom side, carefully bring a small vial, test tube, microtube, or Pasteur pipette in contact with the target hanging drop. Capillary action will force the hanging drop and its content to transfer into your vessel.
11. How do I set up my liquid handling system to perform automated pipetting?
Liquid handling systems allow you to quickly scale up your experiments in Perfecta3D™ Hanging Drop Plates. If you own or have access to a Biomek FX liquid handling robot, please refer to our guides here. Methods for other liquid handling systems will be added soon.
12. What is the intellectual property status of your products?
All products have patents pending. Additionally, the Perfecta3D™ Cell Culture Scaffolds have issued patents. We are actively developing new technologies to expand the application and utility of our products. Please check back often for updates.
13. How do I set up my plate reader to acquire measurements directly from a Perfecta3D™ Hanging Drop Plate?
Many fluorescence- and colorimetric-based assays (e.g. alamarBlue assay) can be conveniently performed directly on the Perfecta3D™ Hanging Drop Plate, without having need to transfer the contents to another microplate. To measure the florescence or absorbance of the assay mixtures, simply place the Perfecta3D™ Hanging Drop Plate into a plate reader and acquire the data as you normally would. Set your plate reader to read from the bottom of the plate. Most plate readers have the option to automatically adjust the Z-focus, which can improve the quality of measurements. The A1 position (notched corner) is located at 12.13 mm (X), 8.99 mm (Y). The well-to-well distance is 4.5 mm.
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