In situ Hybridization of Embryos with Antisense RNA Probes - Caenorhabditis Elegans: Molecular Genetics and Development - page 254

Fig. 2 (A) Appearance of worms in Hypaque meglumine in 1.5-mL Eppendorf tubes. The tube on the
left has been inverted and worms are suspended throughout. After 5 min at room temperature, or after a
brief 2000 rpm centrifugation, the worms will float to the top (right tube) and can be removed by pipetting
the top layer. (B) 80 m L droplet of worms on a glass slide. (For color version of this figure, the reader is
referred to the web version of this topic.)
Prepare a hydration series of methanol:DEPC-ddH 2 O dilutions in five clean
50-mL Coplin jars as follows:
#1 - methanol pre-chilled at 20 C
#2 - 90% methanol at room temperature
#3 - 70% methanol at room temperature
#4 - 50% methanol at room temperature
#5 - DEPC-ddH 2 O at room temperature
Prepare 50 mL of NTF in a Coplin jar and warm it to 37 C in an incubator.
Place an aluminum disc ( Fig. 3 ) onto crushed dry ice for at least 5 min and bring the
disc, still on dry ice, near a dissecting microscope. Make sure the block is smooth and
free from surface irregularities, to allowmicroscope slides to make good contact with it.
Fig. 3 Aluminum disc, approximately 125 mm in diameter 15 mm, on dry ice inside a Styrofoam
shipping carton.