Akura™ 384 Spheroid Microplate vs. Akura™ 384 ImagePro
Both Akura™ 384 Plates have black walls to minimize cross-talk and a unique well design for near-complete medium exchange without spheroid loss.
The Akura™ 384 Spheroid Microplate has a 125 µm thin Polystyrene membrane. The plate is especially suited for high throughput applications, lytic and biochemical assays, and basic confocal imaging endpoints.
The Akura™ 384 ImagePro has a flat, ultrathin, 25 µm gas-permeable membrane made of FEP (fluorinated ethylene propylene) to minimize RI mismatch and it is compatible with high NA objectives.
If you require a high-resolution, high-content imaging endpoint, we recommend the Akura™ 384 ImagePro Plate.
Overview of Akura™ 384 Spheroid Microplate
High throughput screening applications – Confocal imaging – Bio-chemical endpoints
The ultimate spheroid screening plate unifies robust liquid and spheroid handling with automated high-throughput screening applications. The flat bottom and black walls of the InSphero 384 well plate facilitate confocal imaging, while the special well design 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 are preserved in long-term culture
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High-content imaging quality with a continuous, flat-bottom, 125 μm Polystyrene membrane, and a black-walled body
- Improved spheroid/organoid formation with the Akura™ Tilting Stand by keeping the plate at 30 degrees during cell aggregation
Cat.-No.: CS-PC24
Features of Akura™ 384 Spheroid Microplate
10-pack of individually wrapped Akura™ 384 Spheroid Microplates with black-walled polystyrene body bonded to transparent, continuous, 125 µm membrane, ultra-low-attachment coated, sterile, includes lid.
Architecture
- The unique well design of the SureXchange™ ledge protects the spheroid by leaving a minimal amount of residual volume (2-3 μl)
- ANSI/SLAS standard format for full compatibility with automated liquid handling and high-content imaging systems
- A two-component plate consisting of a black polystyrene body and a 125 μm transparent bottom membrane
Handling
- >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
Imaging
- Continuous flat bottom with excellent planarity for imaging applications with immersion objectives, minimal optical aberration, and constant spheroid z-position
- The black-walled body eliminates fluorescent crosstalk between wells
- Defined 1-mm quick-reference region of interest for increased imaging speed
Resources
At InSphero we use our Akura™ Spheroid Microplates 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
- 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
Akura™ 384 Spheroid Microplate FAQs
Q: Why do you recommend pre-wetting the wells prior to spheroid seeding?
A: Pre-wetting the wells of the Akura™ 384 Plate is recommended prior to seeding to prevent the 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.
Please 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 takes the Akura™ Plate from the incubator and centrifuges 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: Could you recommend a cell concentration for my cell suspension for generating spheroids?
A: For long-term growth profiling, we recommend starting with low cell numbers (250 – 500 cells per well of 50 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.
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 centrifuging the Akura™ 384 Plate after cell seeding?
A: We recommend briefly centrifuging 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 it in a microplate centrifuge for 2 minutes at 250 RCF. Afterward, 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 plate's 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 volume (> 50 μl). This will lead to an almost complete removal of the medium, with a consistent residual medium volume.
Add 50 μl of the 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 125 μm thin membrane.
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 the 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 practices 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 the 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 the low-frequency medium exchange.