Specimen enrichment for optical tomography analysis

A system for enhancing gating performance of a cell sorter to prepare an enriched specimen for optical tomography cell analysis, the system comprising:

• one or more processors;
• and
• a memory device storing a set of instructions that, when executed by the one or more processors, causes the one or more processors to:
• a) introducing a controlled specimen including a plurality of known objects treated with an Ab cocktail into a fluorescence-activated cell sorter (FACS) to generate a first set of 2D event data, wherein the known objects include a plurality of known target objects;
• b) generating a first scatterplot of the first set of 2D event data;
• c) locating a first set of the known target objects in the first scatterplot, wherein the known target objects each meet or exceed a target size;
• d) constructing a first gate boundary within the first scatterplot to produce a first gate around a portion of the first set;
• e) counting the known target objects within the first gate to produce a first value;
• f) comparing the first value to a first predetermined value, if the first value meets or exceeds the first predetermined value then proceeding to the next operation g), otherwise, adjusting the first gate boundary and repeating operations e) through f) until the first predetermined value is satisfied;
• g) generating a second scatterplot containing a subset of the first set of 2D event data in the first scatterplot;
• h) constructing a second gate boundary around a second set of known target objects in the second scatterplot to produce a second gate;
• i) counting the second set of known target objects within the second gate boundary to produce a second count;
• and
• j) comparing the second count to a second predetermined value, and if the percent of target objects within the second gate meets or exceeds a second predetermined value the first and second gate boundaries are stored in memory operation otherwise, the second boundary is adjusted and operations i) and j) are repeated until the predetermined criteria is met for the second gate.