384 METHODS AND FORMULAS DS 21.11 



21.11 Klaatsch test. 1896 Kahlden and Laurent Kahlden and Laurent 1896, 178 



REAGENTS REQUIRED: A. wator 100, hematoxylin 1, picric acid \; B. 1% acetic acid 

 method: [sections] — > A, 1-2 mins. -^ B, 30 sees. -^ balsam, via usual reagents 

 result: cartilage, blue; bone, yellow. 



21.11 Kolliker test. 1928 Schmorl Schmorl 1928, 263 



reagents required: A. acetic acid; B. sat. aq. sol. indigo-carmine 

 method: [sections of decalcified bone] -^ A, till transparent —> B, 15-60 sees. -^ wash, 

 till differentiated -^ balsam, via usual reagents 



21.11 von Korff 1907 1780, 69:515 



REAGENTS REQUIRED: A, Water 100, glycerol 7, acid fuchsin 2, orange G 1 



method: [sections of chrome, or dichromate, fixed materials]—^ water—* A, 1 min. -^ 



95% ale., till differentiated -^ balsam, via usual reagents 

 RECOMMENDED FOR: histogenesis of bone and teeth. 



result: osteoblasts and odontoblasts, orange; fibrils in degenerating cartilage, red; 

 other tissues, yellow. 



21.11 Lundvall 1927a 766,62:353 



REAGENTS REQUIRED: A. \% ammonia; B. 4% formaldehyde; C. 0.02% alizarin red S in 

 90% ale. 



method: [embryos, or small vertebrates, fixed and bleached in AF 31.1 Lundvall 1927] 

 -^ A, 24 hrs. or till all acid neutralized — > B, 24 hrs. or till all ammonia removed — * 

 95% ale, till formaldehyde removed —> C, 1-2 days — > 95% ale, till no more color 

 comes away — > [l^enzyl benzoate, via abs. ale. and benzene for museum mounts] or 

 -^ balsam, via usual reagents, for microscope slides 



recommended for: demonstration of bone and calcified structures in wholemounts. 



21.11 Lundvall 1927b 766, 62:353 



STOCK solutions: L 0.1%, toluidine blue in 95% ale. IL water 30, 95% ale. 69, acetic 

 acid 1, alizarin red S 0.02 



reagents required: A. 1% ammonia; J5. 4% formaldehyde; C. water 15, 95% ale. 35, 

 acetic acid 0.5, stock I 10, stock II 40; D. 0.01% acetic acid 



method: [embryos, or small vertebrates, fixed and bleached in AF 31.1 Lundvall 1927] 

 -> A, 24 hrs., or till all acid neutralized -^ B, 24 hrs., or tUl all ammonia removed -^ 

 A, 1 day, 40°C. —> D, till no more color comes away -^ 70% ale, till no more color 

 comes away ^^ 95% ale. thorough wash — > [benzyl benzoate, via abs. ale. and ben- 

 zene, for museum specimens] or — » balsam, via usual reagents, for microscope slides 



recommended for: demonstration of both cartilage (blue) and bone (red) in whole- 

 mounts. 



21.11 Morpugo 1908 test. 1928 Schmorl Schmorl 1928, 269 



reagents required: A. sat. sol. lithium carbonate; B. 0.025% thionin 100, ammonia 



0.1; C. sat. sol. phosphotungstic acid; D. sat. sol. picric acid 

 method: [celloidin sections of F 7000.0000 Mliller 1859 fixed, and nitric acid decalcified, 

 bone] -^ water, thorough wash -^ A, 1 min. -^ B, 3-5 mins. -^ wash -^ C, 5 mins. -^ 

 D, 2-3 mins. — > rinse — » 95% ale. — > balsam, via usual reagents 



21.11 Nollister 1934 20540b (abstr. 1935), 10:37 



stock solution: water 96.5, glycerol 13, acetic acid 3.5, chloral hydrate 0.8, alizarin 



red S to sat. 

 reagents required: A. 1 to 4% potassium hydroxide; B. 1-4% potassium hydroxide 



100, stock 0.1 

 method: [whole fish]-* A, till translucent-^ B, till bones stained —> A + increasing 



quantities of glycerol under UV light till bleached and clear -* pure glycerol 



21.11 Rait 1935 11139b, 19:80 



formula: 70% ale. 100, 1% acetic acid 1, sat. sol. alizarin in abs. ale. 5 

 method: [material, fixed and stored in 10% formaldehyde, adjusted with sodium 

 borate to pH 9] -^ running water, 24 hrs. -* 70% ale, till saturated -^ stain, 12-24 

 hrs. -* 70% ale, wash — ♦ neutral mountant, via usual reagents 



