Open-dish incubator for live cell imaging with an inverted microscope.

Area of Science:

• Biotechnology
• Microscopy
• Cell Biology

Background:

• Live cell imaging requires stable environmental conditions, particularly temperature and focal stability.
• Standard inverted microscopes often lack integrated incubation solutions, hindering long-term live cell studies.
• Existing solutions can be expensive or complex to implement.

Purpose of the Study:

• To design and fabricate an inexpensive, stable cell culture incubator for inverted light microscopes.
• To provide a reliable platform for live cell imaging applications, including those requiring instrument insertion.
• To develop a cost-effective alternative to commercial incubators for microscopy.

Main Methods:

• Fabrication of a temperature-controlled aluminum ring mini-incubator (ringcubator).
• Integration of the ringcubator onto the stage of an inverted light microscope.
• Testing of temperature stability and focal stability with oil-immersion lenses.
• Development of two versions for culture dishes and coverslips.

Main Results:

• The ringcubator maintains cell culture temperature at 37°C with high stability.
• Achieved focal stability for 20-25 minutes using oil-immersion lenses after thermal equilibrium.
• Both versions cost less than $400 to fabricate.
• Assembly requires simple wiring and approximately 3 hours.

Conclusions:

• The developed ringcubator is a cost-effective and efficient solution for live cell imaging on inverted microscopes.
• It offers excellent temperature and focal stability, crucial for demanding live cell experiments.
• Particularly suitable for techniques like electrophysiology and micromanipulation requiring instrument access to the sample.