Insphero Akura 384 ImagePro User manual

Akura™ 384 ImagePro
Quick Start Guide
Thank you for choosing InSphero's Akura™ 384 ImagePro for your 3D cell culture experiments.
This Quick Start Guide contains important information to get you started immediately. For detailed
instructions please refer to the Product Manual on shop.insphero.com.
Akura™ 384 Plate Components
A. Akura™ 384 well plate
B. Transparent lid
Advantages of the Akura™ 384 ImagePro
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Figure 1. Akura™ 384 Plate components.
4. 1 mm diameter flat bottom observation chamber enables
simple spheroid/organoid localization, observation and ROI
identification.
5. Akura™ 384 Plate is compatible with state-of-the-art
imaging and automated liquid handling systems enabling HTS
applications.
1. Convenient scaffold-free formation of 3D cell models via
cellular self-assembly in ultra-low attachment (ULA-treated)
plates.
2. The continuous, highly transparent, 25µm fluorinated ethylene
propylene (FEP) membrane results in enhanced imaging
quality and depth, and the black-walled body eliminates
fluorescent crosstalk between wells.
3. SureXchange™ tapered ledge and culture chamber facilitates
easy medium exchange and prevents spheroid/organoid loss
during long-term spheroid growth and analysis.

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Schlieren, Switzerland
InSphero Inc.
Brunswick, ME, USA
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AG. 3D InSight, Akura and InFloat
are trademarks of InSphero AG.
For life science research only. Not
for use in diagnostic procedures.
Generating 3D Cell Culture Models Like Spheroids or Organoids
Important: In order to prevent inclusion of air bubbles, is is recommended to pre-wet the wells of the Akura™
384 Plate. Apply 40μl of cell line medium containing FCS or BSA to each well by placing the tip near to, but not
touching, the bottom of the well.
1. Cell seeding: Count the cells and prepare a cell suspension for seeding, using a final volume per well
of 50µl. For a long-term growth profiling start with low cell numbers (250 – 500 cells per well). If
non-proliferating cells or rapid production of larger spheroids are required, then start with higher
numbers (2,500 cells or more per well). You may wish to try several different concentrations to define
your optimal range.
Important: Ensure a homogeneous distribution of the cell suspension by gently pipetting up and down prior to
seeding. Also, gently add 50µl of cell suspension (≤10 µl/sec) by placing the pipette tips near to, but not touching,
the bottom of the wells.
2. Sedimentation spin: It is recommended to briefly centrifuge the plate for 2 minutes at 250 RCF to
remove air bubbles.
3. Tilt the plate in the incubator to approximately 30° to improve the maturation process.
4. Incubate the plate in a humidified CO2incubator at 37°C. Spheroid maturation typically occurs within
2-5 days of seeding depending on the cell type and culture conditions.
Medium Exchange in the Akura™ 384 ImagePro
1. Place the pipette tip at the ledge of the well (Fig. 2).
2. Remove the medium at low pipetting speed (<30 µl/sec) by aspirating an excess of volume. A minimal
volume of ~2-3µl medium will remain in the well.
3. Add 50µl of fresh medium by placing the pipette tip at the ledge. Use a dispensing rate <50 µl/sec.
4. Place the lid on the Akura™ 384 Plate and place it in a humidified CO2incubator at 37°C.
Figure 2. Spheroid formation in the Akura™ 384 Plate.
InSphero Akura™ 384 ImagePro
For detailed information,
please refer to the Akura™ 384
ImagePro Product Manual.