358
Related Techniques
Comments
If the hemolymph can be obtained aseptically, filter sterilization is not
necessary.
Results
A suspension of free virions is obtained.
Subprotocol 2
Preparation of Inoculum from Whole Bodies or Tissues
If the virus-infected insects are too small to enable collection of
hemolymph, inoculum can be prepared by homogenizing whole bodies
or some tissues.
Materials
Equipment - Dissecting microscope
- Dissecting tray
- Scissors
- Forceps
- Centrifuge and centrifuge tubes
- Homogenizer
- Maximov slides
- Membrane filter
Solutions - Physiological solution
- Culture medium
Animals - Infected insects
Procedure
Dissection 1. Fix the insects on a dissecting tray filled with physiological solution,
and excise some tissue in which the virus is supposed to replicate (e.g.,
fat bodies).
2. Collect excised tissues in a Maximov slide filled with culture medium.
,mogenization 1. Transfer the tissues, with culture medium, into a homogenizer, and
homogenate, or, in the case of very small insects from which sufficient
amount of virion cannot be obtained, homogenate a sufficient number of whole insect bodies in culture medium.
2. Transfer the homogenate into a centrifuge tube, and centrifuge at
1,500 g for 15 min.