Recover live herpes virus from infected mouse ganglia with Minilys
Instrument: Minilys homogenizer
Sample: Mouse trigeminal ganglia
Protocol:
- Precellys kits: 03961-1-203 (CK14 - 0.5mL tubes).
- HSV-2 virus stock with known titer.
- Mouse trigeminal ganglia (TG) harvested three days after HSV-2 cornea infection.
- Medium HBSS with 1 % FBS.
- Virus stock was diluted to desired concentration and 500 μl was transferred to a homogenizer tube.
- A fresh TG was placed in a homogenizer tube with 500 μl of medium.
- All above samples were kept on ice unless specified.
- Samples were homogenized at RT with settings as indicated. Homogenization was paused every 20 seconds and tubes were held on ice for 1 minute then continue homogenization.
Results:
Live virus in homogenates was detected by titration on a Vero cell monolayer in 6-well-plates following standard plaque assay protocol.
The mouse TG tissue required more than 3 k rpm x 45 sec or 4 k rpm x 20 sec to be adequately homogenized (results not shown).
Diluted virus stock (cell free virus) can withstand the homogenization up to 5 k rpm x 60 sec (Fig. 1).
Live virus can be recovered from HSV-2 infected mouse TG.
