High throughput – Secure handling – Superior imaging
The ultimate spheroid screening plate unifying robust liquid and spheroid handling with automated high-resolution imaging. The flat bottom and black walls of the InSphero 384 well plate facilitates high-content imaging while the lower cavity of the wells allows medium exchange, dosing and endpoint determination in the same plate saving time and resources.
- Unique SureXchange™ ledge engineered to prevent unintended spheroid/organoid aspiration
- Spheroid/organoid morphology and functionality is preserved in long-term culture
Superior high content imaging quality with a continuous, flat-bottom, 25 μm, gas-permeable membrane, and a black-walled body
- Improved spheroid/organoid formation with the Akura™ Tilting Stand by keeping plate at 30 degrees during cell aggregation
10-pack of individually wrapped Akura™ 384 Spheroid Microplates with black-walled polystyrene body bonded to transparent, continuous, 25 µm gas-permeable membrane, ultra-low-attachment coated, sterile, includes lid.
- Unique well design With SureXchange™ ledge protecting the spheroid from aspiration while leaving a minimal residual volume (2-3 µl)
- ANSI/SLAS standard format for full compatibility with automated liquid handling and high-content imaging systems
- Two-component plate consisting of a black polystyrene body and a 25 µm, continuous gas-permeable transparent bottom membrane
- >90% medium removal without spheroid loss using manual and automated pipetting
- Ultra-low attachment surface with 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
- Continuous flat bottom with excellent planarity for imaging applications with immersion objectives, minimal optical aberration and constant spheroid z-position
- Ultra-thin bottom improving imaging z-depth and resolution by minimizing light scatter
- Black-walled body eliminates fluorescent crosstalk between wells
- Defined 1-mm quick-reference region of interest for increased imaging speed
At InSphero we are using our Akura™ Spheroid Microplates daily to produce and assay 3D microtissues for our partners in pharma and biotech. Our manuals and protocols contain valuable information and hands-on advice to make sure that these great plates work in your hands perfectly.
- Product Manual Akura™ 384 Spheroid Microplate
- Technical Specifications Akura™ 384 Spheroid Microplate
- Quick Start Guide Akura™ 384 Spheroid Microplate
- [Technical Protocol] 3D Aggregation of Tumor Spheroids in the Akura™ 384 Spheroid Microplate
- [Poster] 3D High-content imaging in Akura™ 384 with minimal optical aberration enables accurate quantitation of co-localized fluorescent signals within the nuclei of a 3D spheroid model
- [Poster] Streamlining 3D high content-imaging by combining automated 3D spheroid culturing, staining and imaging in a multifunctional well-plate
- Certificate of Compliance
Publication for your reference
- Wardwell-Swanson, Judith, et al. “A Framework for Optimizing High-Content Imaging of 3D Models for Drug Discovery.” SLAS DISCOVERY: Advancing the Science of Drug Discovery 25.7 (2020):709-722
Q: Why do you recommend pre-wetting of the wells prior to spheroid seeding?
A: Pre-wetting the wells of the Akura™ 384 Plate is recommended prior to seeding to prevent inclusion of air-bubbles. For pre-wetting, apply 50 µl of PBS to each well by placing the tips placing near to, but not touching the bottom of the well.
Centrifuge the Akura™ Plate for 2 minutes at 250 RCF and incubate it in a humidified CO2 incubator for at least 1 day. Before cell seeding take the Akura™ Plate from the incubator, centrifuge the Akura™ Plate for 2 minutes at 250 RCF. Aspirate the PBS by placing the tip at the ledge of the upper cavity of the well. Aspirate until the PBS is removed from each well. A small amount of PBS (< 2-3 μl) remains in the bottom of the chamber.
Q: Why is it not allowed to touch the bottom of the well?
The Akura™ 384 Plate consists of a 25 μm gas-permeable membrane. Therefore, never allow the pipette tip to touch the bottom of the well. There is a risk of accidentally puncturing this ultra-thin membrane with a pipette tip.
Q: Could you recommend a cell concentration for my cell suspension for the generating of spheroids?
A: For long-term growth profiling, we recommend to start with low cell numbers (250 – 500 cells per well of 50 µl). If use of non-proliferating cells or rapid production of larger spheroids are required, start with higher numbers (from 2500+ cells per 70 µl). Generally, we recommend to try different concentrations for defining your optimal range when using new cell types.
Q: What is the optimal volume per well in the Akura™ 384 Plate?
A: To achieve optimal conditions, gently deliver 50 µl (pipetting speed < 10 µl/sec) of cell suspension into each well of the Akura™ 384 Plate by placing the pipette tips far into the wells (avoid touching the well bottom).
Important – To obtain spheroids with uniform size and cell composition it is essential to assure a homogeneous distribution of the cells by gently pipetting up and down prior to seeding into the Akura™ 384 Plate.
Q: Why do you recommend to centrifuge the Akura™ 384 Plate after cell seeding?
A: 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 in order to promote cell-aggregation and spheroid formation.
For that, place the lid on the plate and spin in a microplate centrifuge for 2 minutes at 250 RCF. Afterwards, incubate the plate in a humidified CO2 incubator at 37 °C for 2-5 days.
Q: How do I exchange the medium in the Akura™ 384 Plate without disturbing or losing the spheroids?
A: To prevent spheroid loss during the exchange of media, place multi-channel pipette tips at the ledge by slowly sliding down along the inside of the well wall (angled slightly toward the top of the plate) until a subtle resistance can be felt.
Note: Proper aspiration with a multi-channel pipette is possible only row-wise. Carefully and slowly remove the medium by aspirating an excess of volume (> 50 μL). This will lead to an almost complete removal of the medium, with a consistent residual medium volume.
Add 50 μL of pre-warmed medium by placing the pipette tip at the ledge of the plate well and gently dispense at a slow pipetting speed. Never allow the pipette tip to touch the bottom of the well as it consists of a 25 μm gas-permeable membrane. There is a risk of accidentally puncturing this ultra-thin membrane with a pipette tip.
Important - when using automated liquid handling devices, an off-center alignment of the vertical pipette tip will achieve the same effect.
Q: What is the best way to prevent evaporation in the outer wells of my plates?
A: 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:
- 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).
- For culture in the Akura™ 384 Plate, at least 40-50 µl of medium in each well is recommended. Medium exchange frequency can also be increased to every other day or daily if conditions dictate.
- We recommend the use of the InSphero Incubox to reduce edge effects when performing long-term culture with low-frequency medium exchange. The Incubox will be launched in Q4/2021 and will be available on shop.insphero.com.