High-throughput assessment of anemophilous pollen size and variability using imaging cytometry

Area of Science:

• Botany
• Ecology
• Genetics

Background:

• Pollen grain size influences plant community structure, dispersal, and species identification.
• Limited pollen size data, due to traditional measurement constraints, restricts understanding of its variability.
• Existing methods often measure fewer than 50 pollen grains per species, hindering broad ecological insights.

Purpose of the Study:

• To address the data gap in pollen size information.
• To develop and validate a high-throughput method for pollen size assessment.
• To investigate pollen size variability in relation to plant phylogeny and ecological factors.

Main Methods:

• Utilized imaging flow cytometry (IFC) for high-throughput pollen analysis.
• Measured over 500,000 individual pollen grains from 100 anemophilous species (2018-2022).
• Developed a workflow for high-throughput data analysis and validated IFC estimates against literature data.

Main Results:

• IFC provides high-throughput, statistically robust pollen size measurements.
• Demonstrated agreement between IFC-derived pollen size estimates and existing literature values.
• Assessed pollen size variability within the context of plant phylogeny.

Conclusions:

• IFC overcomes sampling effort and throughput limitations in pollen size measurement.
• This approach enables robust ecological research on pollen size at large temporal and spatial scales.
• Facilitates answering broad ecological questions previously constrained by data availability.