Immortalized osteosarcoma cell line using the Leica Stellaris 8 TauSTED

Live Cell Imaging of Immortalised Osteosarcoma Cell Line

Cellbox Solutions | University of Bielefeld, Medical School EWL

The limits of light microscopy have been overcome by breakthrough technologies in the last decade. Several research groups around the world have developed new methods to improve the resolution for the identification of cellular events, particularly in the area of live cell imaging. However, super-resolution microscopes are a very expensive and complex type of laboratory equipment that not every lab can afford. In addition, it can take several months or years to master the operation of the instrument, making it even more difficult to utilize. This report describes the use of a

Cellbox to transport U-2OS cells from a laboratory at the Medical Faculty OWL, University Bielefeld to a demonstration facility in Mannheim.

"All planned imaging experiments

were successful and the cells were

examined under the microscope as

confluent, viable and with vibrant activity."

THE PROJECT

The advantage of live cell imaging is the visualization of transient and dynamic cellular processes avoiding fixation artifacts. This technology is especially critical for the study of signaling pathways and molecular interactions.In this project, U-2OS cells (an immortalized os teosarcoma cell line) stably expressing different genetic constructs were seeded on day one at 5x103 cells/ml in ibidi 8-well μ-slides at 300 μl per well and in 35 mm dishes (MatTek) at 2 ml per dish using DMEM without phenol red supplemented with 10% fetal calf serum and antibiotics. Cells were grown for two days. The Cellbox was packed with confluent U-2OS cells at 37°C and 5% CO2 on day three. Cells were kept in the Cellbox during the cab ride between the University of Bielefeld and the Bielefeld central station, the subsequent train ride from Bielefeld central station to Mannheim central station, and the cab ride between Mannheim central station and a local hotel for an overnight stay.

- June, 2023