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Updated: Sep 17, 2025
07:22
Basophil Activation Test for Allergy Diagnosis
Published on: May 31, 2021
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A hand-operated microfluidic sample preparation-to-analysis workflow for simplifying the basophil activation test.
Nicolas Castaño1, Kaiser Chua1, An Nguyen1
1Department of Mechanical Engineering, Stanford University, USA. sindy@stanford.edu.
Lab on a Chip
|July 2, 2025
Summary
A new microfluidic device simplifies basophil activation testing (BAT) by enabling sample preparation and stabilization, making this allergy diagnostic more accessible. This innovation extends analysis time, reducing the need for immediate processing and specialized labs.
Area of Science:
- Allergy diagnostics
- Immunology
- Microfluidics
Background:
- Basophil activation testing (BAT) is crucial for allergy diagnosis but limited by requirements for fresh blood, specialized equipment, and technical expertise.
- Current BAT protocols necessitate timely sample processing, hindering broader clinical application.
Purpose of the Study:
- To develop and validate a microfluidic sample preparation device (μF-prep) to simplify BAT.
- To enhance accessibility of BAT by overcoming limitations of fresh blood analysis and specialized laboratory needs.
- To stabilize basophil samples for extended analysis windows using flow cytometry.
Main Methods:
- A hand-operated microfluidic device (μF-prep) was developed for parallel basophil stimulation and staining.
- Barcoded staining allowed sample pooling for stabilization in lyse/fix buffer.
- An XGBoost-enabled pipeline automated data analysis, unpooling conditions and generating activation metrics.
- μF-prep performance was compared to conventional BAT preparation (conv-prep) using whole blood from peanut-allergic and non-allergic donors.
- Sample stability was assessed after storage in lyse/fix buffer for up to 7 days at 4 °C.
Main Results:
- The μF-prep device required <2 minutes of active user engagement for sample preparation.
- The automated analysis pipeline demonstrated excellent agreement with manual gating.
- μF-prep showed comparable dose-response activation levels, maximum activation, AUC, and EC50 values to conv-prep.
- Basophil activation levels remained stable in samples stored in lyse/fix buffer for 7 days at 4 °C compared to day 0 analysis.
Conclusions:
- The μF-prep device significantly simplifies sample preparation for BAT.
- Sample stabilization extends the analysis window, removing the need for immediate processing and overnight shipping.
- This technology has the potential to broaden access to BAT for allergy diagnostics.
