Materials and Reagents

1. Standard cell line [e.g. MeWo (ATCC^® HTB-65 ^TM); ARPE19 (ATCC^® CRL-2302^TM); Vero (ATCC^® CCL-81 ^TM)]
   
2. Fluorescently labeled virus stock (genetically engineered VZV-66GFP, HSV1-gC-GFP and PRV-IE180-GFP were kindly provided by Prof. Paul R. Kinchington, University of Pittsburgh, Pittsburgh PA)
   
3. Growth medium for the cells listed above (e.g. item #6, Fibroblast medium)
   Note: However, it can be any other medium for other pairs of virus and cells.
   
4. 4% paraformaldehyde (EMS 15710) in phosphate buffered saline with divalent cations (e.g. Biological Industries, catalog number: 020201A )
   
5. High glucose DMEM (Biological Industries, catalog number: 010551A )
   
6. Fetal calf serum (Biological Industries, catalog number: 041271A )
   
7. Glutamine (Biological Industries, catalog number: 030201 )
   
8. Penicillin-streptomycin (Biological Industries, catalog number: 030311 )
   
9. Agarose low EEO (Hispanagar, catalog number: D1500 )
   
10. Fibroblast medium (see Recipes)
    
11. Overlaying media for free virus (see Recipes)
    
12. Crystal violet (Sigma-Aldrich, catalog number: C0775 ) (see Recipes)
    

Equipment

1. 24-well tissue culture plates
   
2. Incubator 37 °C, 5% CO₂
   
3. Epifluorescent inverted microscope
   

Procedure