172 Connective Tissue (chap. 13) 



8. Transfer to slide with a drop of solution. 



9. Dry thoroughly; do not add more solution. 



10. When completely dry, add moiniting medium and cover glass. 



comments: 



To bring out density and distribution of the mineral in undecalcified 

 bone, see Frost (1959). He describes methods with basic fuchsin, silver 

 nitrate, alizarine red S and others. 



Dowding (19'>9) uses methyl methacrylate as a plastic embedding 

 solution, and grinds down the bone as thin as 30 microns. 



Hause {1959) describes a block for holding the bone while sawing 

 it and recommends a Razor Sa\v blade #35-ST (Lipshaiv Manufac- 

 turing Company) as a good cutting instrument. 



Kropp {19'^4) describes a plastic embedded method using heat and 

 pressure. 



Yaeger (1958) uses freeze-drying and vacuum infiltration with butyl 

 methacrylate-ethyl methacrylate. 



Norris and Jenkins (1960) describe a method using epoxy resin for 

 preparation of bone for radioautography. The resolution is good; 

 there are no chemicals in resin to produce artifacts in nuclear emul- 

 sions; the medium does not warp or chip when machined or abraded. 

 (Polyesters and methacrylate do distort Avhen machined.) The design 

 of a metal and Incite mold for the embedding is included. Sections 

 can be made with a microtome (6-10 microns), with a circular saw 

 (50-100 microns) and groinrd to give sections as thin as 10-50 mi- 

 crons. 



Alizarine Red S Method for Embryos (Bone Formation) 



fixation: a hardening action. 



Hollister (193-1), 70% alcohol for fish, removal of scales desirable: 

 several days. 



Hood and Neill (1948), 95% alcohol, organism preferably free of hair 

 or feathers: 3 days. 



Richmond and Bennett (1938), 95% alcohol: 2 weeks. 



Some specimens require decolorization. The best method is to lay 

 specimen in 95% alcohol in white tray. Place in direct sunlight for 

 24 hours each side. An Alpine sunlamp was used by Hollister on sun- 

 less days. 



