Assurance of clonality

After seeding single cells into 96 well plates, cells will be allowed to settle to the bottom of the well (typically several hours) or can be centrifuged.

Cells are then imaged by the Cell Metric CLD which is a dedicated whole well imager for assurance of clonality. Using the 10 plate stacker on the Cell Metric CLD, one cassette of 96 well plates should provide more than enough clones to identify a knock-out clone.

Imaging is carried out at Day 0 and subsequent days of growth (using confluence) and ultimately colony formation. This can be 10-21 days depending on cell types.

A library of images is created for each well and it is possible to track back the history from the colony all the way back to Day 0 and confirm if it started from 1 cell or not (see images below).

Cell Metric Colony

Cell Metric 2 Cells

Cell Metric 1 Cells

Using Next Generation DNA sequencing (NGS) and bar-coded primers, it is possible to pool a sample of cells from the wells rather than process them independently. Once the correct mutation has been sequenced and identified, it can be easily deconvoluted and tracked back to which well/plate this clone came from.

A report can be rapidly generated for the well history of the top clones. This documentation package is called the Clonality Report and is importantly something which customers can use to support their clonality claims.

To find out more

Next Steps