Introduction
In this application, the combination of the CQ1 with the stocker with Incubator and CellPathfinder enabled continuous and automatic long-term observation of the process of single cells forming colonies.
The use of cells that need to be cultured for a longer period of time, e.g. iPS and ES cells to induce differentiation and primary cell cultures, has become increasingly important.
Single cell cloning, which isolates a single clonal (monoclonal) population, is also an important method for cell line establishment. However, colonies generated by limiting dilution methods sometimes merge with other colonies until they reach a recoverable cell number.
These experiments require a lot of manpower to check the status of the cells every day, which is a burden on the experimenter.
With the use of an automated solution continuous observation during long-term culture can be automated, reducing the experimenter's workload.
Experiment
Materials
- Cell Line: 293 (HEK) GFP
- Plate: Greiner #655090
Imaging and analysis
- Magnification: 4x Dry lens
- Excitation wave length: 488nm, Bright Field
- Field of View: 4 field of view per well
- Time lapse: every 12 hours for 8 Days
- Analysis: Measure cell area by CellPathfinder.
Result
In this application, the combination of the CQ1 with the stocker with Incubator and CellPathfinder enabled continuous and automatic long-term observation of the process of single cells forming colonies.
In addition, by returning the plates to the incubator when not in use for observation, the imager is no longer occupied for long periods of time, making it possible to conduct live cell experiments for extended periods of time.
Left: Time-lapse movie of colony formation
Right: Cell area magnification change over time
Cell areas detected by CellPathfinder: Above is Day 0 and below is Day 8. Day 0 is a gradation from 0 % to 100 %. Day 8 gradates from white to red between 0 and 50 % (everything above 50 % is red).
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