Histology - Acrylic resin
Embryos set in acrylic resin were serially sectioned at 2 micron thickness.
Protocols
Embryo collection and preparation
The embryo was prepared for sectioning as described in Embryo collection - Acrylic resin
Sectioning
The block was clamped in a vice type ultratome chuck and the block face trimmed with a glass knife on an ultratome. The glass knife was replaced by a diamond Histoknife with a built in trough. 2 micron sections were then cut onto distilled water and collected in strings of 15 sections (2 strings per slide) directly onto glass slides. The sections were allowed to stretch out on the water for 2 mins at 70oC on a hot plate. The water was drained away onto tissue paper and the slides allowed to dry vertically at room temperature overnight. The sections were then sealed onto the slide by heating on the 70oc hot plate for 30 mins.
Staining
The sections were stained with Solochrome Cyanin stains A and B as below
Solution A in a coplin jar in a water bath at 65oC for 5 mins. Washed in tap water and allowed to 'blue' in tap water for 2 mins. Rinsed in distilled water then placed in a coplin jar containing Solution B in a water bath at 65oC for 10 mins. Washed in tap water, rinsed in distilled water and allowed to air dry vertically at room temperature.
Slides mounted in DPX applied directly to the slide and the coverslip carefully placed on top.
Image Capture
After staining the slides were available for image capture.
