instawell 0.3.0.dev3

Nice tools for plotting and analyzing thermal shift assay wells

InstaWell

Tools for organizing, processing, and visualizing thermal shift assay (TSA) data.

Why?

"Man I hate copying and pasting stuff in excel... I wish I could very quickly get from raw TSA data + layout to dose-response curves and Tm values without a million clicks."

Features

• Flexible layouts → parse arbitrary condition fields (e.g., concentration | ligand | protein | buffer)
• Long/wide transforms and replicate averaging
• Background subtraction using a non-protein control (NPC) marker
• Min-max scaling and derivative-based min-temperature (Tm-like) extraction
• Prism-style 4PL fit (log10 domain with logEC50) + diagnostics CSV
• Plotly figures: raw per-well, processed averages, and min-temperature scatter
• Jupyter widgets to browse generated figures interactively

Install

pip install instawell
# Optional: notebook extras (recommended)
pip install 'instawell[notebook]'

Python ≥3.10 recommended, as well as the tool uv for development environments.

Or install from source:

git clone https://github.com/DavidHein96/InstaWell.git
cd InstaWell
uv sync

What does this do?

Instawell takes your instrument's TSA output (temperatures × wells) and a layout that describes each well's condition (e.g., concentration|ligand|protein|buffer). It then organizes, QC-plots, and computes dose-response summaries.

1) Inputs

A) TSA output (raw instrument CSV) Each column after the temperature is a well (A1, A2, …). Values are the signal (e.g., fluorescence).

Temperature,A1,A2,A3,B1,B2,B3
25.0,  120,115,118,  130,126,127
26.0,  118,114,117,  128,125,126
...

B) Layout (maps well positions to conditions) This ties each well to a condition string your pipeline understands.

well_row,1,2,3
A, 0|DMSO|NPC|PBS,  1.5|DrugX|ProteinA|PBS,  6|DrugX|ProteinA|PBS
B, 0|DMSO|NPC|PBS,  1.5|DrugX|ProteinA|PBS,  6|DrugX|ProteinA|PBS

• well_row + column number → well name (e.g., A1, A2, …).
• Condition fields (order matters): concentration | ligand | protein | buffer
• NPC in protein = non-protein control for background.

---

2) What the pipeline does (high level)

1. Ingest & organize

   • Melts the wide TSA matrix into long format (one measurement per row).

   • Expands condition fields into columns and builds:

     • unqcond = concentration|ligand|protein|buffer
     • well_unqcond = well|concentration|ligand|protein|buffer

   • 📄 01_raw_organized_data.csv

   Example (long rows):

   Temperature,well,value,concentration,ligand,protein,buffer,unqcond,well_unqcond
   25.0,A2,115,1.5,DrugX,ProteinA,PBS,1.5|DrugX|ProteinA|PBS,A2|1.5|DrugX|ProteinA|PBS
   26.0,A2,114,1.5,DrugX,ProteinA,PBS,1.5|DrugX|ProteinA|PBS,A2|1.5|DrugX|ProteinA|PBS
   ...
   

2. (Optional) Filter bad wells

   • You inspect raw per-well plots to flag odd traces (bubbles, spikes, drifts).
   • 📊 raw_figures_widget(...) helps you browse quickly.
   • 📄 02_filtered_organized_data.csv (even if nothing removed, for traceability)

3. Average replicates

   • Averages value over wells sharing the same unqcond at each temperature.
   • Produces long + pivoted wide tables keyed by Temperature.
   • 📄 03_averaged_data.csv, 03_averaged_data_long.csv

4. Background subtraction (NPC)

   • For each (ligand, buffer, concentration) with protein ≠ NPC, subtracts the matching NPC column.
   • Leaves NPC columns out of the final set (less heavy-handed removal).
   • 📄 04_bg_subtracted_data.csv, 04_bg_subtracted_data_long.csv

5. Min–max scaling (QC convenience)

   • Scales each unqcond trace to [0, 1] to make shapes comparable.
   • 📄 05_min_max_scaled_data.csv, 05_min_max_scaled_data_long.csv

6. Derivative & min temperature (“Tm-like”)

   • Computes derivative curves and finds the temperature at the minimum derivative per unqcond.
   • 📄 06_derivative_data(_long).csv
   • 📄 07_min_temperatures.csv (has concentration, ligand, protein, buffer, min_temperature)

7. Dose–response (Prism-style 4PL)

   • Fits a 4-parameter logistic in log10 dose space using logEC50 (zeros are excluded there).
   • Outputs parameter table (Bottom, Top, logEC50, EC50, Hill, SEs, 95% CIs, RSS/RMSE, AIC/BIC) and point-wise diagnostics.
   • 📄 08_curve_params.csv, 08_curve_diagnostics.csv

Quick start

Also see the example notebook

from instawell import (
    setup_experiment,          # Step 00 - creates a folder structure and saves metadata
    ingest_data,               # Step 01 - organizes raw data and extracts conditions from the layout
    filter_wells,              # Step 02 - filtering of wells (required to be run)
    average_across_replicates, # Step 03 - groups replicate wells
    subtract_background,       # Step 04 - NPC background subtraction
    min_max_scale,             # Step 05 - min-max scaling
    calculate_derivative,      # Step 06 - derivative computation 
    find_min_temperature,      # Step 07 - min-temperature extraction
    calculate_curve_params,    # Step 08 - 4PL curve fitting
    load_experiment_context,   # load existing experiment context from disk
)

exp = setup_experiment(
    experiment_name="demo1",
    experiments_root="experiments/demo1",
    raw_data_path="data/demo1/raw.csv",
    layout_data_path="data/demo1/layout.csv",
    condition_fields=("concentration","ligand","protein","buffer"),
    condition_separator="|",
    empty_condition_placeholder="0",
    non_protein_control_marker="NPC",
)

# --- Pipeline (pass along the exp) ---
ingest_data(exp)                  # -> 01_raw_organized_data.csv
filter_wells(exp)                 # -> 02_filtered_organized_data.csv
average_accross_replicates(exp)   # -> 03_averaged_data.csv (+ long for easier formatting)
subtract_background(exp)          # -> 04_bg_subtracted_data.csv (+ long)
min_max_scale(exp)                # -> 05_min_max_scaled_data.csv (+ long)
calculate_derivative(exp)         # -> 06_derivative_data.csv (+ long)
find_min_temperature(exp)         # -> 07_min_temperatures.csv
calculate_curve_params(exp)       # -> 08 curves: params/diagnostics CSVs

Jupyter widgets (requires jupyter notebook)

from instawell import raw_figures_widget, processed_figures_widget, min_temp_figures_widget

# Raw per-well plots (discrete colors per well, only for raw and filtered data)
raw_figures_widget(exp)

# Processed/averaged plots (everything after averaging step)
processed_figures_widget(exp, data_source="bg_subtracted", color_scale="Thermal")

# Min-temperature scatter (with selectable modes in generator args)
min_temp_figures_widget(exp, mode="log10_fit", color_scale="Viridis")

The widget wrappers preserve the docstrings & signatures of the underlying generators. Use show_help=True to display the docstring in a collapsible panel. You can also save individual figures from the generators by passing save_figs=True to the widget, since they are in plotly they are saved as HTML files by default, which you can open in a browser or convert to PNG using kaleido.

Typical files (by step)

experiments/<name>/

01_raw_organized_data.csv
02_filtered_organized_data.csv
03_averaged_data.csv
03_averaged_data_long.csv
04_bg_subtracted_data.csv
04_bg_subtracted_data_long.csv
05_min_max_scaled_data.csv
05_min_max_scaled_data_long.csv
06_derivative_data.csv
06_derivative_data_long.csv
07_min_temperatures.csv
08_curve_params.csv
08_curve_diagnostics.csv
experiment.log                  # A log file of pipeline steps
01_raw_plots/...                # saved HTML
03_averaged_plots/...
experiment_info.json            # Parsed layout and wells metadata
experiment.json                 # ExperimentContext configuration
filtered_wells.txt              # List of excluded wells for extra reference
original_raw_data.csv          # Copy of the original raw data
original_layout_data.csv       # Copy of the original layout data

Key concepts

• Condition fields - configured via ExperimentContext.condition_fields and joined with condition_separator to derive:

  • unqcond (unique condition string)
  • well_unqcond (unique well + condition)

• NPC background subtraction - subtracts a matching non-protein control column per panel; leaves NPC traces out of the final set.

Configuration (ExperimentContext)

Common fields:

• experiment_dir: output directory (CSV, plots, logs)
• raw_data_path, layout_data_path, temperature_column
• condition_fields: tuple of fields (order matters)
• condition_separator: string used to join/split fields (e.g., "|")
• empty_condition_placeholder: placeholder for 'blank' conditions. A fully blank well would be e.g., "0|0|0|0"
• non_protein_control_marker: e.g., "NPC"
• log_to_file, log_level: pipeline logging

Development Notes

This tool is currently best suited for use from a jupyter notebook, but a CLI and Dash app are planned. The dash app will be very helpful as it will greatly simplify creating a layout file from plate maps.

Another TODO is to fully finish implementing the fuzz testing suite for the various data processing functions.

License

This project is licensed under the GNU AFFERO GENERAL PUBLIC LICENSE Version 3 (AGPL-3.0)