Principles & Protocols

Necessary Supplies

• 0.5% Formvar in Chloroform solution.
• 50% glycerol in water solution.
• Methanol
• Chloroform
• grids
• Newspaper
• carbon coater
• conical beaker
• probe sonicator
• dipping jar
• crystallizing dish

Technique

• Make 0.5% formvar solution
  • Dissolve 1 g formvar
  • into 200 mL Chloroform
  • Allow to dissolve overnight

Making holey film

• Place 50 mL of formvar solution in a conical beaker
• add 20 drops of 50% glycerol solution
  • more drops equal larger holes
• swill contents of conical beaker
• add contents to dipping jar
• Probe sonicate dipping jar contents on maximum power for 1 min
  • contents should be warm and milky in color
  • contents should be used to make grid within 1/2 hour of sonication
• Dip a clean microscope slide into the solution and set to dry staning up on newspaper.
  • microsope slides will keep for years

Applying film to grids

• Score grid to release film
• float the film on water in a crystallizing dish.
• Place grids on film
• Pick up film and grids with newspaper.
• Allow the paper to dry.
• Dissolve holes with methanol for 10mins.
  • Make sure to avoid covering top of grids with methanol
  • A petri dish with methanol whetted filer paper works well.
• Allow to dry thoroughly.
• Evaporate with carbon.
• Dissolve plastic away with chloroform for 10 minutes
  • Use a petri dish with whetted filter paper again
• Evaporate another layer of carbon.

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 05 Oct 2006