Topographic imaging of spores in different relative humidity levels is performed in tapping mode using a Multimode AFM (Bruker, Inc.) housed within a separate scanner-mounted ECU. Humidity is adjusted as described for the ECU used in relaxation kinetic measurements with an identical sensor mounted <1 cm from the sample. 750 nm glass beads (Corpuscular Inc., Cold Spring, NY) are mixed 1:1 with the spore suspension for absolute size-change reference and to account for any piezo-induced artefacts of the high humidity. The sample is dispersed onto a mica substrate (Ted Pella Inc., Redwood, CA) for imaging. Germinated, or otherwise unresponsive, spore shells are excluded from analysis. The image is processed for plane fit and maximal cross-sectional height relative to the substrate is obtained using the Nanoscope software package (Bruker, Inc.). Heights of different spores are normalized using the average height of all spores. A separate set of measurements is performed to determine the maximum height change of spores. For these measurements, spore height is measured under 3% RH, 50%, and when spores are immersed in water. The measurement at 50% RH is used to calibrate these measurements against the first set of measurements, therefore height measurement at 50% is not used in the analysis of height vs RH in Fig. 3b .
Nanoscope software package
Manufactured by Bruker
Sourced in United States
The Nanoscope software package is a comprehensive data acquisition and analysis software suite designed for Bruker's atomic force microscopes (AFMs). It provides intuitive tools for imaging, spectroscopy, and advanced analysis of nanoscale samples.
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2 protocols using nanoscope software package
1
Spore Topographic Imaging at Various Humidity
2
Quantifying Palm Oil-Butter Particle Surface Roughness
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The AFM analysis was performed to quantify the surface roughness of the palm oil-butter particles. The same samples used for SEM imaging were used for AFM analyses. AFM experiments were conducted using a Bruker Dimension Icon AFM with ScanAsyst automatic image optimization mode based on PeakForce Tapping (Santa Barbara, CA, USA). ScanAsyst-Air probe (k ~0.4 N m -1 , tip radius < 10 nm) (Bruker Corporation, Santa Barbara, CA, USA) was employed in all AFM measurements. The oscillation frequency was 2 kHz. All images were recorded in air and under atmospheric conditions (22°C, 25% RH). The scanning field was kept at 10 μm by 10 μm for all the particle samples. Two dimension (2D), three dimension (3D), and roughness data were acquired from the Nanoscope software package (Bruker Corporation, Santa Barbara, CA, USA). The morphology of the surface sample was characterized by the arithmetic mean surface roughness (R a ):
is the height of the center plane, and Z i is the ith height sample out of n total samples, as Z i tends toward Z ep surface roughness decreases. n is the number of samples within a given area.
is the height of the center plane, and Z i is the ith height sample out of n total samples, as Z i tends toward Z ep surface roughness decreases. n is the number of samples within a given area.
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