Biopharma Insights Here, we present a protocol to quantify allergen-loaded particles by flow cytometry. Ambient particulate matter particles may act as carriers of adsorbed allergens. We show here that flow cytometry, a method widely used to characterize suspended solids >0.5 µm in diameter, can be used to measure these allergen-loaded particles.
Total time
~3-4 hours per sample
Steps
1
Collect PM10 particulate matter samples
Use a high-volume sampler to collect ambient PM10 particles onto filters over a defined period.
▶ 00:49
2
Stain particles with fluorescently labeled antibody
Incubate extracted particles with a fluorescent antibody specific to Bet v 1 to label allergen-bound particles.
▶ 02:54
3
Analyze stained particles by flow cytometry
Use flow cytometer to detect and quantify particles with Bet v 1-bound fluorescence.
▶ 06:04
4
Compare allergen-positive particle counts across seasons
Evaluate flow cytometric data to observe variations in allergen-loaded particles between high and low pollen seasons.
▶ 08:29