Individual curves

Results from plots

Table of all results

help notes

►Calibrator tab

►Load calibrator data and select a well: csv, txt or xlsx file types are accepted
►Columns of calibrators or data can be selected over a range from 'start' to 'end' or on different Excel sheets
►A curve (magenta -) is fitted to the calibrator fluorescence data (blue +) and intial rate calculated (red O)
►The number of points selected for fitting should be chosen to cover the fluorescence range in the sample data
►It is advisable to limit the number of points to be fitted to avoid noisy, unused data late in the time course
►A polynomial correction algorithm is applied to correct the data for substrate depletion and inner filter effect (green X)
►The quality of the fit is shown in the graphs to the right
►Sample data are loaded using the dialogue box as csv, txt or xlsx
►Figures of fluorescence versus time are shown and can be arranged by number of rows
►'No correction' or 'polynomial correction' can be applied using the radio buttons

►Plots and Curve tabs

►Transformed data plots showing thrombin concentration are shown in these tabs
►Results tables are also shown for a selected parameter ('Plots') or all parameters for a single curves ('Curve')
►The end of the lag phase is signalled by a change of 10% in trombin concentration but this can be adjusted
►Tail smoothing is used to select a steady rate to calculate the final Thrombin-alpha-2M contribution (gold curves)
►Smooth the curves after the signal has returned to baseline and smoothing can be seen using the radio button selection
►Graphs can display fluorescence or thrombin concentration with or without subtraction of the Thrombin-alpha-2M activity
►Lag time and ttTail are the points where the curve reaches or returns to 10% (or selected value) of max
►These points are also used as boundaries to calculate the area under the curve

►Other tabs

►'Results' presents a table of results
►'Raw data' shows all the raw data and fluorescence. Thrombin or corrected thrombin values (with or without alpha-2-M-thrombin) can be selected
►'Calibrator data' shows raw calibrator data columns as selected
►'Settings' summarises the setting used to generate the analysis as an aid to reproducibility
►'Help' includes a theme selector drop down to permit control of fonts, colour and design

Please cite this reference on reproducible analysis in publications

Longstaff C, Development of a Shiny app tool to simplify and standardize the analysis of hemostasis assay data: communication from the SSC of the ISTH, J Thromb Haemost, 15: 1044-6, 2017 DOI 10.1111/jth.13656

A publication is available where the app has been tested on difficult data

Jackson, J et al, Sources of bias and limitations of thrombinography: inner filter effect and substrate depletion at the edge of failure algorithm, Thrombosis Journal, 21: 104, 2023 doi.org/10.1186/s12959-023-00549-5

Please contact me with problems or suggestions relating to:

Thrombin generation assays app 0.992bx4 last accessed 2025-09-05 at drclongstaff@gmail.com

More detailed help notes and the code can be found on my github site: here
Other apps and links for reproducible analysis in haemostasis assays are available here and please check out the Youtube channel of help videos