Akura™ 96 Spheroid Microplate
The Akura™ 96 Spheroid Microplate is the super convenient culturing plate for your daily spheroid/organoid culturing in every lab environment, from manual to automated handling. The plate is designed for lossless spheroid/organoid formation and handling as well as reliable cell culturing and analysis.
- All-in-one plate allowing spheroid formation, long-term culturing and analysis in one single plate
- Unique SureXchange™ ledge engineered to prevent unintended spheroid/organoid aspiration and to retain consistent minimal residual volume (5 -7 µl) during medium exchange
- Spheroid/organoid morphology and functionality are preserved in long-term cell culture by unique ULA coating
- Enhanced imaging speed and resolution through flat bottom well
- A conical well design with SureXchange™ ledge protecting the spheroid/organoid from aspiration while leaving a minimal residual volume (5-7 µl).
- ANSI/SLAS standard format for full compatibility with automated liquid handling systems
- A leak-free and glue-free transparent COP 96-well plate
- >90% medium removal without spheroid loss using manual and automated pipetting
- Ultra-low attachment surface with a stable cell-repellent coating
- Biologically inert, and non-degradable surface compatible with cell culture and in-plate post-culture processing steps such as cell lysis and fixation procedures
- The flat bottom well minimizes optical aberration and ensures constant spheroid z-position
- Defined 1-mm quick-reference region of interest for increased imaging speed
- Cyclo-olefin polymer (COP) plate material with 0.8 mm bottom thickness for good imaging quality
Product Documents
Frequently Asked Questions
A detailed protocol for the production of spheroids in the Akura™96 Spheroid Microplate is provided in the product manual. Below are answers to some frequently asked questions to help get you started.
What are the main characteristics of the Akura™ 96 Plate?
Optical properties:
- COP (Cyclo-Olefin Polymer, 92% transparency 400-800 nm) as plate material
- Thin well bottom of 0.8 mm
- Low skirt height of 0.4 mm. High NA objectives (e.g., 20X and 40X) may be used to image the outer wells of the plate
Automation-friendly:
- Excellent planarity across plate (below 80 μm) for reliable spheroid transfer and precise medium exchange
Reduced evaporation:
- Optimized distance (200 μm) between the customized low-evaporation lid and plate reduces evaporation in outer and edge wells
Standard SLAS plate height:
- 14.35 mm plate height
- Maximum volume 280 μl
Why do you recommend pre-wetting of the wells prior to cell seeding?
Pre-wetting the wells of the Akura™ 96 Plate is recommended prior to seeding to prevent inclusion of air bubbles. For that, apply 40 μl of your cell medium to each well by placing the tips far into the wells. Remove the pre-wetting solution by placing the tip at the ledge of the upper cavity of the well. Aspirate medium until it is completely removed from each well. A negligible amount (< 5-7 μl) may remain in the bottom of the chamber well.
Could you recommend a cell concentration for my cell suspension for the generating of spheroids/organoids?
For long-term growth profiling, we recommend starting with low cell numbers (250 – 500 cells per well of 70 μl). If the use of non-proliferating cells or rapid production of larger spheroids is required, start with higher numbers (from 2500+ cells per 70 μl). Generally, we recommend trying different concentrations for defining your optimal range when using new cell types.
What is the optimal volume per well in the Akura™ 96 Spheroid Microplate?
To achieve optimal conditions, gently deliver 70 μl (pipetting speed < 10 μl/sec) of cell suspension into each well of the Akura™ 96 Plate by placing the pipette tips near, but not touching, the bottom of the wells.
Important: For spheroids with uniform size and cell composition, it is essential to ensure a homogeneous distribution of the cells by gently pipetting up and down prior to seeding into the Akura™ 96 Plate.
Why do you recommend centrifuging and tilting the Akura™ 96 Spheroid Microplate after cell seeding?
We recommend to briefly centrifuge the plate after cell seeding to remove any air bubbles and to force the cells to the bottom of the well to promote cell-aggregation and spheroid formation.
For that, place the lid on the plate and spin in a microtiter-plate centrifuge for 2 minutes at 250 RCF. Afterwards, incubate the plate in a humidified CO2 incubator at 37 °C for 2-5 days.
Tilt the plate in the incubator to approximately 30° or use Akura™ Tilting Stand (InSphero, CS-AG11) to improve the maturation process.
How do I exchange the medium in the Akura™ 96 Spheroid Microplate without disturbing or losing the spheroids?
To prevent spheroid/organoid loss during the exchange of media, the SureXchange™ ledge at the inside wall of each well serves as an anchoring point for the pipette tip. Just place the tip at the ledge of the well and remove the medium at low pipetting speed (>30 μl/sec). A minimal volume of ~5-7 μl will remain in the well.
Then, add 70 μl of fresh medium by placing the pipette tip at the ledge, using a dispensing rate of <50 μl/sec.
Important: When using automated liquid handling devices, an off-center alignment of the vertical pipette tip will achieve the same effect.
What is the best way to prevent evaporation in the outer wells of my plates?
Evaporation in the outer (perimeter) rows of wells is a phenomenon common to most low-volume culture platforms, and thus requires careful attention to maintaining proper humidity control. If not controlled, pronounced evaporation can result in concentration or precipitation of media components (serum, salt) that can impact spheroid formation or health, and can alter the effective concentration of a compound/additive in the medium over the course of a long-term experiment.
To provide maximum humidity control when using the Akura™ Plates, we recommend the following:
1. Use an incubator with good humidity control (>95% of rel. humidity), and exercise best practice in maintaining and minimizing loss of humidity (e.g., minimize incubator door opening and closing).
2. For culture in the Akura™ 96 Spheroid Microplate, at least 50-70 μl of medium in each well is recommended and can be increased to a maximum of 80 μl if incubator humidity control is a persistent issue. Medium exchange frequency can also be increased to every other day or daily if conditions dictate.
3. We recommend the use of the InSphero Incubox™ (CS-AH11) to reduce edge effects when performing long-term culture with low-frequency medium exchange. The InSphero Incubox™ is available on shop.insphero.com.
Which clearing reagents can you recommend?
Several well-established clearing reagents are available, such as ScaleS, which has shown very good performance with our spheroid models. In general, we strongly recommend testing any clearing reagent in Akura™ Plates prior to routine use to confirm compatibility, especially if it includes organic solvents. Some clearing reagents can be relatively aggressive and, in certain cases, may interact with or damage the plate material. Conducting a preliminary compatibility test helps ensure optimal performance and prevents potential issues.
