Cell number
When using cell lysis extracts, the analyte concentrations can be normalized to the cell number and the extraction solvent volume used. The functionality of the Cell normalization tool is described by an example.
Cell number and Cell extraction volume can be defined in LIMS > Sample registration > Tissue and cell factors in the corresponding fields. In this manner, each sample extract’s analyte concentration (canalyte) can be normalized by the Cell extraction volume (Vextr) and the Cell number (Ncells) according to equation:
ccells = (canalyte ∙ Vextr) / Ncells
Example
| Sample | Cell number | Cell extration volume [µL] |
|---|---|---|
| Sample 001 | 1300400 | 50 |
| Sample 002 | 5700100 | 50 |
| Sample 003 | 800600 | 50 |
For Sample 001 and the exemplary canalyte = 1.23 µM this will be:
ccells (Sample 001) = (1.23 µM ∙ 50 µL) / 1300400 cells = 47.29 ∙ 10-18 mol / cell
The cell number of the normalized amount of substance (ncells) can be considered (e.g. ncells per 106 cell).
ccells (Sample 001) = 47.29 (10-18 mol) / cell = 47.29 pmol / 106 cell
ccells (Sample 002) = 10.79 (10-18 mol) / cell = 10.79 pmol / 106 cell
ccells (Sample 003) = 76.81 (10-18 mol) / cell = 76.81 pmol / 106 cell
The Cell Normalization Tool does not affect the analyte concentration unit in the WebIDQ database (which is µmol/L by default).
Subsequent tissue factor normalization:
For subsequent data normalization after data has already been processed in WebIDQ, please select the corresponding plate-run in LIMS and Delete measurements. Afterwards enter the cell factor values and re-import your corresponding data files again.