FLVP assays
Perform preparation of FLVPs as described in Viral and bacterial isolates, propagation and preparation of stocks.
Prepare a 10-fold dilution series of the culture or natural sample to be enumerated so that ~105 cells mL–1 of the target (FLVP-specific species) will be obtained.
Perform all subsequent steps under subdued light, since the stain will fade if exposed.
Add 0.1 mL FLVP working stock to 0.9 mL sample in a microcentrifuge tube for each slide to be prepared and vortex to mix.
Allow up to 30 min for adsorption of FLVPs to target cells.
Filter each 1 mL sample onto a 0.2-µm Anodisc filter using a 0.45-µm HA filter for backing.
Pipette 10 µL mounting medium onto the surface of a slide.
Place the filter over the drop.
Pipette 10 µL mounting medium onto the surface of the filter.
Place a 25 × 25 mm coverslip over the filter.
Observe the slides under blue-light excitation and count the cells with a fluorescent halo.
