From: Advances in rare cell isolation: an optimization and evaluation study
Parameter | Optimization goal | Definition | Dependencies |
---|---|---|---|
Limit of detection (LOD) | 0.24 cells/mL peripheral blood | Detection of at least one cell of interest in 4.2Â mL blood or 2.5e7 total cells | Test sensitivity, recovery, Ltotal |
Enrichment factor | >500 | Ratio between the cell-of-interest frequency before and after enrichment | Depletion factor and recovery |
Depletion factor | >625 | Ratio between the leukocyte amount before and after enrichment | Bead amount high |
Recovery | 80% | Measure of target cell loss | Bead amount low |
Initial leukocyte amount, Ltotal | 2.5 × 107 | Assay start with leukocyte cell count | Blood volume 3–7 mL, enrichment factor, cell of interest frequency, |
Residual leukocytes, Lfinal | 2 × 104 to 4 × 104 cells | Leftover leukocytes after depletion | Depletion factor |
Assay time | 2Â h | Complete assay time including isolation and analysis | RBC lysis quality and enrichment factor |