JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:24 min • August 31st, 2026
Take tubes containing viral capsids, which are virus-like particles without genetic material.
Heat the tubes at a constant temperature for varying durations.
A shorter heating duration mildly stresses the capsid proteins, while prolonged heating disrupts the capsid and causes aggregation.
Rapidly cool the tubes to prevent further structural changes.
Transfer the liquid onto a paraffin film.
Place a transmission electron microscopy (TEM) grid onto the droplet. The capsids attach to the grid's surface.
Blot the grid with a filter paper to remove excess fluid.
Wash with a buffer to remove loosely attached capsids, then blot the grid again.
Dip in uranyl acetate, a heavy metal stain that binds to the capsid surface, making them appear darker under TEM.
Blot the grid and place it on a plate. Store the plate in a desiccator to remove moisture.
Image the grid using TEM to visualize heat-induced capsid damage.
Perform heat treatment as before except substitute PBS for 10 millimolar HEPES, pH 7.4 and do not further dilute capsids after treatment. Next, pipette the entire 15 microliter drop of treated capsid solution onto paraffin film. Grab the grid edge with tweezers allowing them to close on the edg
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