Dehydration: 1. Two changes of for 30 minutes each. 70% ethanol 2. and 70% ethanol (2:1) mixture: slowly add one part of 70% ethanol (drop by drop) to L.R. White two parts of L.R. White, and shake gently (otherwise the mixture will become milky). Incubate specimen in the mixture for 1 hour. 3. Incubate specimen in two or three changes of pure L.R. White for 1 hour each, or overnight at room temperature (specimen can be stored in L.R. White at 4 C for weeks if necessary). Embedding: 1. specimen in Place specimen in bottom of the capsule and fill up with Embed Gelatin Capsules. L.R. White to the brim. 2. in 60 C oven for 24-48 hours. Polymerize Sectioning and Staining: 1. Cut ultrathin sections and on formvar-carbon and allow it air-dry mount coated nickel grids overnight. 3. Stain with for 15 minutes and for 1-2 minutes. uranyl acetate lead citrate Observation: 1. Observe Transmission Electron Microscopy 2. Take pictures Grids fro mounting ultrathin sections:
Dehydration: 1. Two changes of 70% ethanol for 30 minutes each. 2. L.R. White and 70% ethanol (2:1) mixture: slowly add one part of 70% ethanol (drop by drop) to two parts of L.R. White, and shake gently (otherwise the mixture will become milky). Incubate specimen in the mixture for 1 hour. 3. Incubate specimen in two or three changes of pure L.R. White for 1 hour each, or overnight at room temperature (specimen can be stored in L.R. White at 4 C for weeks if necessary). Embedding: 1. Embed specimen in Gelatin Capsules. Place specimen in bottom of the capsule and fill up with L.R. White to the brim. 2. Polymerize in 60 C oven for 24-48 hours. Sectioning and Staining: 1. Cut ultrathin sections and mount on formvar-carbon coated nickel grids and allow it air-dry overnight. 3. Stain with uranyl acetate for 15 minutes and lead citrate for 1-2 minutes. Observation: 1. Observe Transmission Electron Microscopy 2. Take pictures Grids fro mounting ultrathin sections: