SEM Imaging of Fungal Spore Germination

Three to six capsules were harvested from controls and each treatment, air-dried, opened on stubs to disperse the spores, and processed according to Rabbi et al. (2020) (link). Germinated spores were fixed per the TEM protocol and critical point dried rather than being infiltrated with resin. The specimens were sputter-coated with 50 nm of Au/Pd using a Denton Desk II Vacuum Sputter Coater and imaged using a scanning electron microscope (SEM) (QUANTA FEG 450; FEI) with 5 kV acceleration voltage.

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Renzaglia K.S., Ashton N.W, & Suh D.Y. (2023). Sporogenesis in Physcomitrium patens: Intergenerational collaboration and the development of the spore wall and aperture. Frontiers in Cell and Developmental Biology, 11, 1165293.

Publication 2023

Desk II Vacuum Sputter Coater FEG 450

Acceleration Capsules Rabbi Resin Scanning electron microscope Spores Vacuum

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Variable analysis

independent variables

Treatment

dependent variables

Germinated spores

control variables

Number of capsules harvested (3 to 6)
Air-drying of capsules
Dispersing spores on stubs
Processing method as per Rabbi et al. (2020)
Fixation of germinated spores for TEM protocol
Critical point drying instead of resin infiltration
Sputter-coating with 50 nm of Au/Pd
Imaging using SEM with 5 kV acceleration voltage

controls

Controls (unspecified number of capsules)

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