The group led by Dr. Yoshimura in the Ozawa Laboratory at the University of Tokyo is performing fluorescence live cell imaging, especially single molecule imaging in living cells. However, photobleaching of fluorescent signal is a significant problem in live cell imaging. In order to observe weak fluorescence signal in real time, strong excitation is required, which inevitably leads to a dimmer image.

In order to reduce fluorescence fading, there is a method to perform live cell imaging in a redox state similar to in-vivo environment and hypoxic condition. A hypoxic environment can be performed by using a commercially available microscope stage-top incubator, but if reagents are added to the sample, these reagents must also be kept in equilibrium with the same hypoxic condition.

Challenges and Solutions

In order to solve this problem, TFS designed a new system (CEIM-C001) that enables all samples and reagents that maintain hypoxic condition on the microscope stage while the experiment is being performed. TFS made this happen by placing the samples and reagents in a stage-top incubator with hypoxic condition while having a flow path, including a pump, into the incubator.
The CEIM-C001 allowed the lab to conduct longer experiments of several minutes at a time, while the conventional experimental setup would only allow 20-30 seconds of observation.

Image courtesy of Ozawa Laboratory, The University of Tokyo

- Links to tubing pumps and piezoelectoric micro pumps used in this device