Clonal Validation & Expansion

Confirmation of clonality is a critical step in the generation of biopharmaceutical cell lines. With superior brightfield image quality, the Celigo can reliably visualize individual cells in a well, even if the cell is adjacent to the well wall, and allows for verification that a clone originates from a single cell. Typically, limiting dilution is performed in multi-well plates which are then repeatedly imaged on the Celigo over a period of time. When colony growth is seen after a few days of culture, it is then possible to retrospectively confirm that it grew out of a single cell. Clonal validation on Celigo provides several benefits:

• Fast brightfield scanning of multi-well plates (384-well plate can be scanned in <5 min)
  
• Accurate image segmentation for detection of single cells in a well
  
• Heatmap representation of the number of cells/well
  
• Growth tracking report for monitoring cell growth over time
  
• Efficient database management of images and data

Clonal validation on Celigo. Whole-well (384-well plate; top) and zoomed brightfield images (bottom) of single CHO-S cells after seeding.

Clonal expansion on Celigo. Zoomed brightfield images of a single CHO-S cell expanding into a colony.

Automatic detection of single CHO-S cells on Celigo. Representative brightfield single cell segmentation (left) and resulting heat map of the number of cells per well (right).

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