Tcs sp8 aobs laser microscope

The reinfection assay was performed as previously described but with slight modification^{10 (link)}. The symbiont F190/B001 was grown for 5–7 days on ISP2 medium plates. GFP-labeled A. faecalis SCSIO B001 was first grown in liquid media (LB with kanamycin) for 2 days. Then, 200 μL of bacterial culture was plated onto the fungal petri dish and cocultivation was carried out for 1 week. The sample was then washed with fresh saline (3 X) and the remaining symbiont visualized using a Leica TCS SP8 AOBS Laser Microscope at 510/520 nm.

According to the methods detailed by Partida-Martinez and Sulemankhil^{8 (link)}, growing mycelia of symbiont F190/B001 (0.5 ml) were transferred to a centrifuge tube containing 0.5 mL 0.85% saline. A 10 µL aliquot of the mixture was placed onto a microscope slide and treated with 0.5 µL Live/Dead BacLight SYTO 9 and an equal amount of propidium iodide (Live/Dead BacLight Bacterial Viability Kit, Molecular Probes-L7007). A 15 min incubation in the dark (RT), ProLong Gold antifade reagent (Molecular Probes–P36935) was employed to enhance fluorescence. The sample was then incubated for another 20 min and analysed using a Leica TCS SP8 AOBS Laser Microscope at 510/520 nm.