Akura™ 384 ImagePro
The Akura™ 384 ImagePro is our high-end solution plate that unifies robust liquid and spheroid handling with automated high-resolution imaging. The flat bottom and black walls of the 384-well plate facilitate high-resolution, high-content imaging, while the special well design allows medium exchange, dosing, and endpoint determination in the same plate, saving time and resources.
- 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 remnant media volumes (< 2-3 µl) during medium exchange
- Spheroid/organoid morphology and functionality are 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
- The unique well design with SureXchange™ ledge protects 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
- A two-component plate consisting of a black polystyrene body and a 25 μm, continuous gas-permeable FEP membrane
- >90% medium removal without spheroid loss using manual or 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
- 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
- The black-walled body eliminates fluorescent crosstalk between wells
- Defined 1-mm quick-reference region of interest for increased imaging speed
Compare Akura™ 384 ImagePro and Akura™ 384 Spheroid Microplate
- 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 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.
- 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.
If your main focus is on biochemical assays and basic imaging, we recommend the Akura™ 384 Spheroid Microplate.
Product Documents
Frequently Asked Questions
A protocol for the production of spheroids in the Akura™ 384 Plate is provided in the product manual. Below are answers to some frequently asked questions to help get you started.
What is the difference between Akura™ 384 Spheroid Microplate and the Akura™ 384 ImagePro Plate?
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 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.
- 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.
If you require a high-resolution, high-content imaging endpoint, we recommend the Akura™ 384 ImagePro Plate.
Why do you recommend pre-wetting of the wells prior to spheroid seeding?
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 near, 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.
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.
Could you recommend a cell concentration for my cell suspension for the generating of spheroids?
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 50 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™ 384 Plate?
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 ensure a homogeneous distribution of the cells by gently pipetting up and down prior to seeding into the Akura™ 384 Plate.
Why do you recommend centrifuging and tilting the Akura™ 384 Plate after cell seeding?
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 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.
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™ 384 Plate without disturbing or losing the spheroids?
The special well design allows medium exchange, dosing and washing without disturbing or losing the spheroids. Medium exchange can be performed using multi-channel pipettes or semi-or automated liquid handling devices.
(a) Use of multi-channel pipettes:
When using multi-channel pipettes, place the tips at the SureXchange™ ledge (Figure 1) 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.
(b) Use of semi-or automated liquid handling devices:
When using automated liquid handling devices, an off-center alignment of the vertical pipette tip is necessary to place the pipette tip on the SureXchange™ ledge, see Figure 2. For more detailed information, please refer to the Product Manual of the Akura™ 384 ImagePro.
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™ 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.
3. We recommend the use of the InSphero Incubox™ (CS-AH11) (Figure 3) to reduce edge effects when performing long-term culture with low-frequency medium exchange. The InSphero Incubox™ is available on our shop.
