BOEDEKER'S METHOD 



49 



BONE 



water 1 change. Tone in 1% gold chlo- 

 ride with 3 drops of glacial acetic acid 

 per 100 cc, 5-10 min. Wash in re- 

 distilled water 1 change. If sections 

 do not have a light purple color place 

 in 2% oxalic acid until the entire section 

 has the slightest blue or purplish tinge. 

 Pour off as soon as tissue gets slightly 

 blue. Remove residual silver salts in 

 5% sodium thiosulfate 5-10 min. Wash, 

 dehydrate, clear and mount. Note : 

 the Coplin jars used must be cleaned in 

 Cleaning Fluid. The Bodian method 

 has been adjusted for the demonstra- 

 tion of melanin by Dublin, W. B., Am. 

 J. Clin. Path., 1943, 7 (Technical Sec- 

 tion), 127. 



Boedeker's Method, see Enamel matrix. 



Bogoroch, see Radioautographic Technique. 



Boling, see Teeth, Decalcification. 



Bollinger Bodies, see Borrel Bodies. 



Bone. A good account of methods is 

 provided by Shipley (McClung, pp. 

 344-352). Examination without decal- 

 cification involves the cutting and 

 grinding of thin sections. The instru- 

 ments used by dentists for the making 

 of sections of undecalcified teeth are of 

 the greatest service and should be pur- 

 chased or borrowed. If they are not 

 available Grieves' method for dental 

 tissues is suggested. In order to de- 

 termine the structure of bone with 

 organic material removed, Shipley ad- 

 vises cutting away all soft parts after 

 which the bone may or may not be split. 

 Place in tap water, or in a 2% aq. gelatin, 

 to which a loop full of culture of B. coli 

 has been added. After 5-6 days wash 

 in running water 24-48 hrs. in a stink 

 cupboard. This will dissolve and wash 

 away all organic material. Sterilize the 

 bone by boiling or immersion in alcohol. 

 Saw into sections, grind these to the 

 necessary thinness and polish. De- 

 hydrate in ether. Dry thoroughly and 

 mount in balsam. Routine examination 

 includes some method of fixation, de- 

 calcification and staining. Hematoxylin 

 and eosin are recommended, likewise 

 phosphomolybdic acid hematoxylin and 

 Mallory's connective tissue stain. 



For different structural components 

 special techniques are reauired. Bone 

 corpuscles may be isolated by putting a 

 thin section of bone in concentrated 

 nitric acid for a few hours to a day. 

 Then place the section on a slide, cover. 

 Pressure on the cover glass will squeeze 

 out ellipsoidal bone cells with their 

 processes (Shipley). Bone lamellae ca,n 

 be peeled off easily when decalcified 

 bone has been allowed to simmer in 

 water for several hours (Shipley). 

 Lacunae and canaliculi. The easiest 

 method is to impregnate sections of 



ground bone with 0.75% aq. silver 

 nitrate for 24 hours. Wash, polish the 

 sections on a fine hone to remove preci- 

 pitated silver, dehydrate in alcohol, 

 clear in xylol and mount in balsam. 

 The lacunae and canaliculi appear black 

 in a yellowish brown background. To 

 impregnate thin sections with acid 

 fuchsin, dry them after extraction with 

 alcohol. Place them in watch glasses 

 in a 20% aq. sol. of acid fuchsin in a 

 desiccator connected with a suction 

 apparatus. Extract air for about an 

 hour and close the dessicator. After 

 24 hrs. the solution will have dried. 

 Rub off ppt. on a hone, pass through 

 xylol and mount in damar or balsam 

 (Shipley). 



Linings of lacunae and canaliculi. 

 (Schmorl's method modified by Ship- 

 ley.) Employ a fixative not containing 

 mercury. Decalcify in Miiller's fluid, 

 wash in running water, embed in cel- 

 loidin and section not over 10 microns. 

 Stain in thionin solution alkalinised 

 by 2 drops ammonia. Transfer with 

 glass needle to sat. aq. phosphotungstic 

 or phosphomolybdic acid. Leave until 

 blue, gray or green. Place in water 

 until sky-blue. Ammonium hydroxide 

 1 cc. and aq. dest. 10 cc, 3-5 min. 

 Several changes 90% alcohol. 95% ale. 

 Clear in carbolxylol and mount in 

 damar (or balsam). This method is 

 suggested for bones of children. 



Processes of young osteoblasts in grow- 

 ing bone. Shipley suggests following 

 treatment of slices of bone of a rickety 

 animal. 4% aq. citric acid 20^30 min. 

 in the dark. Rinse in aq. dest. 1% aq. 

 gold chloride in the dark, 20-30 min. 

 3% formic acid in the dark, 48 hrs. 

 Rinse in aq. dest. and preserve in pure 

 glycerin. Make frozen sections, mount 

 in glycerin and ring with damar, balsam, 

 paraffin or cement. Keep specimens in 

 dark when not is use. 



To determine relative age of deposi- 

 tion the following method has proved 

 useful in senile osteoporosis. Saw sec- 

 tions of bone not more than 0.5 cm. 

 thick and fix in 4% formalin 2-4 days. 

 Decalcify in 6% isotonic formalin, 40 

 cc, 85% formic acid, 60 cc, and sodium 

 citrate, 5 gm. changing every second 

 day for, say, a week, that is until they 

 become flexible and can be penetrated 

 by a fine needle. Embed in celloidin 

 (slow method). Prepare stain by dis- 

 solving 30 gm. potassium alum in 1 liter 

 hot water and by adding 1.5 gm. hema- 

 toxylin crystals. Cool and add 1 gm. 

 chloral hydrate. Ripening in sunlight 

 to rich dark color is hastened by addition 

 of crystal of potassium hydroxide. 

 Stain celloidin sections about 2 days 



