Flow cytometry · one fluorophore per run

MESF Calibrator

Turn measured fluorescence into MESF with the same log–log regression used for the post-COVID EV analysis. Workbooks stay in this browser; only reusable calibrator-lot metadata can be saved to the shared library.

Model log₁₀(MESF) ~ log₁₀(FI)

Choose the workbook

Use one XLSX containing calibration measurements and the values to convert.

XLSX Drop a workbook here or choose a file from this computer

Sample data are processed locally and never uploaded.

Run setup & calibrator lot

Choose the fluorophore yourself; the tool never guesses a channel.

Fluorophore / channel Measurement label (optional)

Calibrator lot

Loading shared library…

What is kept from the original pipeline?

The calculation is deliberately narrow and auditable.

MESF = 0 bead rows are excluded from the fit.
All positive points are fitted by ordinary least squares in log₁₀ space.
Positive sample FI is back-transformed without censoring.
Values outside the measured calibration range are retained and flagged.
R² below 0.995 is highlighted for review, matching the original QC threshold.

Before you calculate

Replicates, negative values & blanks

These rules are applied consistently in the workbook and documented in the PDF report.

Sample replicates stay separateEvery selected value is calibrated individually. For a single technical-replicate summary, calculate the median afterwards in MESF space. Keep biological replicates separate.

Calibration replicates use the medianRepeated bead rows with the same positive MESF assignment are collapsed to their median measured FI before fitting, so that level is not overweighted.

Zero, negative or missing FIThe raw cell is preserved, the MESF result is left blank, and the output is flagged. A logarithmic calibration cannot transform non-positive FI.

No hidden correctionMESF = 0 bead rows are excluded. Blank subtraction, clipping, censoring and LLOQ estimation are not automatic; positive out-of-range values are retained and flagged.

Map the workbook

Select the sheets, header rows, and columns explicitly.

Calibration measurements

Bead curve

run-specific FI

Sheet Header row

Measured fluorescence column MESF assignment column (used for a new/unlisted lot)

Values to transform

Samples

0 columns

Sheet Header row

Filter column names

Ready to calculate?

Raw input values remain unchanged in the downloaded workbook.

MESF Calibrator

Choose the workbook

Run setup & calibrator lot

What is kept from the original pipeline?

Map the workbook

Bead curve

Samples

Curve QC & MESF output

Calibration curve

Points used

First converted values

Add calibrator lot