Insphero Incubox User manual

InSphero Incubox™
Product Manual
PM005, Nov 2021
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Contents
Contents 1
Introduction 2
Key Features 2
Incubox™ Components 3
Step-by-step Protocol 4
Technical Specifications 5

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Introduction
Edge effects due to media evaporation are a common problem in long-term cell cultures,
especially in busy labs with many projects in which incubator doors are frequently opened
and closed. The Incubox™ is a specially engineered container which creates a high-humidity
microclimate inside conventional cell culture incubators and protects your sensitive plates
from large humidity drops and fluctuations –the major cause for evaporation effects.
•Maintain a stable medium volume and accurate compound concentration due to
minimal evaporation over time
•Reduce well-to-well variation by minimizing evaporation-induced reagent
gradients across the plate
•Use all wells of your plate by eliminating edge effects in your culture plate
•Save on plasticware by using up to 38% more wells of your plate
Key Features
1. Incubox™ can accommodate up to 20 microplates and fits on the shelves of a wide range of
incubators.
2. Engineered of aluminum, stainless steel and toughened safety glass (ESG), Incubox™ is
compatible with a wide range of solvents and can be autoclaved in a standard incubator thereby
reducing the risk of contaminations.
3. The Incubox™ is equipped with its own water bath which allows water exchange without
disrupting the incubation conditions.

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4. A defined number of holes on the backside of the Incubox™ ensures gas exchange and maintains
ambient O2and CO2concentrations while also preventing humidity loss.
5. The magnetic closure system allows quick, safe, and one-handed door handling to access the
plates inside the Incubox™.
Incubox™ Components
Figure 1: Photo of the Incubox™ including Akura™ Spheroid Microplates.
The Incubox™ consists of an Aluminum frame and all sides made of toughened safety glass (ESG)
for increased mechanical strength and resistance to changes in temperature. The components on
the inside, the shelf and the water bath are made of stainless steel or materials resistant to heat
and chemicals.
1. Three holes on backside for gas exchange
2. Stainless steel shelf
3. Magnetic doorknob
4. Stainless steel water bath
5. Shelf ledge
*Depth incl. doorknob 375 mm

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Step-by-step Protocol
Please follow the steps below to ensure proper function of the Incubox™ and optimal culturing
conditions:
•Carefully clean all components of the Incubox™ with Ethanol to remove dust.
•Place the Incubox™ directly on a shelf inside your incubator. Make sure it stands stable and
at least 5 cm away from the backside of the incubator. Don’t block the hole on the backside.
We recommend placing the Incubox™on one of the center shelves for easy access of your
plates and to avoid blocking the ventilator on the top of some incubator models.
•Optional: Use the high-temperature sterilization cycle of your incubator to ensure complete
elimination of microbial contaminants.
•Remove the shelf from the Incubox™ and fill water bath with approximately 500 ml of
sterile water including a water bath stabilizer, e.g., Aqua resist. (To re-fill water bath,
remove plates, lift the shelf to one side and re-fill water bath).
•Place the shelf back onto the ledge.
•Close Incubox™ and incubator. Wait at least one hour to ensure stable temperature and high
relative humidity.
•Add your plates and/or culture dishes and start your experiment.

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Technical Specifications
Figures 2 and 3 show the technical dimensions of the Incubox™:
Material:
Incubox™: Aluminum frame; all sides made of toughened safety glass (ESG)
Shelf, water container: Stainless steel
Sterility: Non-sterile
Incubox™ Dimensions:
Box depth: 350 mm (375 mm including door knob)
Box height: 210 mm
Box width: 350 mm
Figure 2: Front view

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Figure 3: Top view

InSphero AG
Schlieren, Switzerland
InSphero Inc.
Brunswick, ME, USA
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3D InSight, Akura, InFloat and Incubox
are trademarks of InSphero AG. For life
science research only. Not for use in
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