Plan apo λ na 1.45 oil objective

Fluorescence images were taken on a Nikon A1 confocal microscope with the Elements software package using a 100× Plan Apo λ (NA 1.45) oil objective. Excitation of fluorescence probes was made with solid‐state lasers and their emission isolated by bandpass filters as follows; SYTO40: 405 nm excitation, 425–475 nm filter, MTG: 488 nm excitation, 500–550 nm filter, FM 1–43: 488 nm excitation, 521.5–554.5 nm filter, FM 4–64: 488 nm or 561 nm excitation and 575–625 nm filter. Short‐term imaging was done on glass slides coated with 0.01% poly‐l‐lysine (Sigma‐Aldrich, St. Louis, MO, USA). Imaging over time was performed with an Attofluor cell chamber (ThermoFisher/Invitrogen, Waltham, MA, USA) using a standard poly‐l‐lysine‐coated coverslip as a base. For long‐term experiments, the sample was kept at a constant growth temperature of 30°C by an Okolab microscope cage incubator. TEM imaging was performed by the Ohio State University Ohio Agricultural Research and Development Center's Molecular and Cellular Imaging Center in Wooster, Ohio.