Load multiple .asyr files and calculate estimated OCR from their raw O₂ sensor readings, then compare treatments across plates. Runs locally in your browser.
Projects
Projects are saved in this browser. Download results for a separate backup before clearing browser data.
1 · Add plates
Your sample has O₂ readings and timestamps, calibration and chamber settings, injection stages, treatment groups, and cell counts. No other file is required.
Plate editor
Click a well to include or exclude it immediately. Drag across wells to select a block for group or cell-count edits; use Shift or Ctrl/⌘ to add to a selection. Striped wells are excluded or instrument flagged. Click a flagged well to include it manually; its instrument flag remains recorded. Wells without a group need an assignment before they can be included. Changes save to the project automatically.
0 wells selected
Choose one well to see its current group and cell count. Background wells cannot be assigned or included. Instrument flagged wells can be included manually.
Normalization values · edit 96-well plate
Click or drag to select cells; Shift-click expands the selection. Use Ctrl/Cmd+C to copy, Ctrl/Cmd+V to paste an Excel range beginning at the selected cell, and Delete to clear selected cells. Double-click a cell to type a value. Only the current plate is changed.
No cells selected
Group colors
2 · Review groups and assay
The calculated rates are raw. Enable normalization when your .asyr has valid cell counts. Background well slopes are subtracted per measurement. Blank and flagged wells are excluded.
Stage assignment uses the .asyr protocol. Your sample has 3 baseline, 3 oligomycin, 2 FCCP, and 3 rotenone/antimycin A measurements. Only combine plates with the same assay type and injection order.
Measurements per stage:
3 · Plate results and selected-plate statistics
Individual plate windows
Open each plate to inspect its well-level graph and values. Edit wells in the plate editor, then recalculate before running combined statistics.
Combine plates for statistics
Choose independent plate replicates below. The one-way ANOVA uses one mean per group per selected plate; multiple wells on a plate are technical replicates.
Respiration profile
Select which OCR metrics appear side by side. Bars and error bars use plate means in the combined view; the Points and Error bars menus also control this graph. Coupling efficiency (%) has its own axis and remains available in the single-metric view.
Single metric
OCR and acidification kinetics
Each point is a group mean from included wells; whiskers show SEM. Injection lines are placed between the protocol stages. Raw pH slope is an acidification signal, not calibrated ECAR.
Plot groups
Hide groups only from plots. ANOVA and exported data still use all included groups. Change order with the arrows.
Plate means
One-way ANOVA (selected plates)
For each metric, the omnibus one-way ANOVA compares group plate means. Pairwise comparisons use the ANOVA residual variance and the correction selected above across all pairs within that metric. The test assumes independent plate-level observations, normally distributed errors, and comparable variance. Groups with fewer than two selected plates are excluded from the test. If the same culture preparation contributes to multiple plates or treatments, a blocked or mixed model may be more appropriate.
Adjusted pairwise comparisons
Well level metrics
Calculation and limits
For each measurement and well, fluorescence is converted to O₂ pressure using the file's F₀ and Ksv: O₂ (mmHg) = (F₀ / corrected emission − 1) / Ksv. A straight line is fitted to O₂ versus time across the stored rate span. Its negative slope is converted to pmol/min using the file's ChamberVolume and COb / CO oxygen concentration factor. Background well mean is subtracted at each measurement. The pH sensor's change in log fluorescence is available as an acidification signal per minute; it is not calibrated ECAR in mpH/min. These are estimates derived from the raw data; Wave's AKOS calculation includes additional processing, and its OCR may differ. Compare against a Wave export before using absolute values interchangeably.