Justification for this choice
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Measurement type chosen based on particles type: particles were very
small, mostly round, and transparent (see example of images below).
VisualSpreadsheet© software version 4.15.1. identifies pixels that
comprise a particle by distinguishing them from the image background
taken automatically at each run. Among other parameters, the software
estimates the Area-Based Diameter (hereafter ABD) and the Equivalent
Spherical Diameter (hereafter ESD). ABD is the diameter calculated based
on a circle with an area equal to the ABD area. The ABD area is
determined by the number of dark pixels in the binary imaged particle,
excluding inner light pixels. ESD is the mean value of 36 feret
measurements. Feret measurements are the perpendicular distance between
parallel tangents touching opposite sides of the particle
(VisualSpreadsheet 5 Particle Analysis Software manual). ABD would be
expected to underestimate the size of particles with internal
transparent components, while ESD would be expected to overestimate the
size of particles with protrusions or flagella (Owen et al.,
2022).
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