Insphero 3D InSight User manual

3D InSight™ Microtissues
Quick Start Guide
Instructions for handling 3D InSight™ Liver and Tumor Microtissues delivered in Akura™ 96 Plates upon
arrival.
Figure 1: On the lower left side, the membrane overlaps the plate. Start with the
lower left corner and remove the seal starting with the first column from left to
right, column by column.
1. Please check the box for any damage that may have occured during transport. If the box is damaged, contact
InSphero customer service.
2. Remove the bag containing Akura™ Plates from the shipping box.
3. Remove temperature logger from shipping box. Verify that green "Recording" LED is blinking intermittently.
Stop the logging process by pressing the "Start" button for 5 seconds (the logger confirms recording-stop by
flashing 10x).
Press the "Start" button again:
a. "Recording" LED flashes green 2x = no temperature exception detected -> proceed with experiments
b. "Status" LED flashes red 2x = temperature exception* detected -> please plug the device into a
computer USB port and send the saved logging files to: [e-mail]
Accepted temperature zone (SZ) = 14°C – 40°C / Higher temperature limit (H1) = 40°C / Lower temperature
limit (L1) = 14°C
4. Wipe the outside of the bag with 70% ethanol and dry. Carefully open the sealed bag and remove the Akura™
Plate containing the microtissues.
5. Do not remove the sealing membrane on the Akura™ Plate as some microtissues may stick to the seal and
need to be recovered by centrifugation.
6. Centrifuge the Akura™ Plate with intact membrane at 250 rcf for 2 minutes at room temperature (each well
contains 70 µl of shipping medium - total weight of plate ~63 g).
7. Place the Akura™ Plate under a microscope to verify each well contains one microtissue. If all microtissues
are present, remove the sealing membrane (Figure 1 and 2). If some wells appear to be empty, please consult
troubleshooting instructions on the backside of this guide or contact InSphero ([email protected])
8. Perform a medium exchange on the tissues according to the instructions in the Technical Protocol (TP007).
9. Allow the microtissues to rest in incubator before starting the experiments. Resting time varies by tissue
type:
For Liver Microtissues - Three hours (minimum)
For Islet Microtissues - Overnight
Figure 2: Hold the plate firmly with your left hand while applying downward
pressure, then remove seal with your right hand until the whole membrane is
released.
www.insphero.comFor technical assistance, please contact InSphero Customer Service.
10. Plan to exchange medium every 2-3 days until you are ready to run assays.

InSphero AG
Schlieren, Switzerland
[phone]
InSphero Inc.
Brunswick, ME, USA
[phone]
Technical Support E-mail
[email protected]
[email protected]
General E-mail and Website
[email protected]
www.insphero.com
InSphero is ISO 9001:2015 certified
All rights reserved, © 2023 InSphero AG. 3D InSight, Akura
and InFloat are trademarks of InSphero AG. For life science
research only. Not for use in diagnostic procedures.
Troubleshooting Instructions
Under certain circumstances, Liver Microtissues can attach to the plate seal or may adhere to the air-liquid
interface due to surface tension interactions. If microtissues appear to be missing after step 7 of this Quick
Start Guide, please follow the steps below to increase the chances for successful recovery of microtissues.
Note: This troubleshooting protocol is not appropriate for all microtissue types. It is not recommended
for 3D InSight™ Islet Microtissues.
a. Centrifuge the Akura™ 96 Plate with seal intact again in a microtiter plate centrifuge at 250 rcf for two
minutes at room temperature.
b. Verify the presence of microtissues at the bottom of the wells using an inverse brightfield microscope.
If any microtissues appear to be still missing, repeat steps a. and b. Persistent failure of complete recovery
may be related to microtissues sticking to the air-liquid interface. In this case, proceed to steps c-f.
c. Place plate under a laminar flow hood, remove lid and gently peel off seal (Figure 1 and 2).
d. Remove 30 µl of medium from the well and re-dispense the same medium from above at a distance of ~2
cm and release 1-2 drops into the well (see illustration below). This will cause any floating microtissue to
sink to the bottom of the well. Alternatively, fast dispense medium directly at the well wall on top of the
microtissue for proper recovery into the well.
e. Verify presence of microtissues at bottom of wells using an inverse brightfield microscope.
f. After successful recovery of the microtissues, proceed with step 8 of this Quick Start Guide.
Aspirate
medium
(~30 µl)
Floating
microtissue
Re-dispense
1-2 drops
~2 cm
3D InSight™ Microtissues Quick Start Guide