STAINS 



490 



STAINS 



I % osmic acid for 24 hours, stain with hematoxylin 

 and differentiate in saturated solution of picric acid. 

 The "granule cells" are l/lue, everything else green. 

 Busch's Mixture, sodium iodid 3 parts, \ c / c osmic 

 acid 100 parts. The iodid is said to enhance the pen- 

 etration of the osmic acid. Biitschli's Method for 

 the demonstration of the foam or alveolar structure of 

 protoplasm : Fix the object with picric acid solution or 

 with iodin-alcohol ; stain by Heidenhain's iron-hema- 

 toxylin method and cut sections from one-half to one 

 micron in thickness. Mount in a medium of low re- 

 fractive power and examine with high magnification. 

 Biitschli-Delafield Hematoxylin, Butschli recom- 

 mends that to a very dilute solution of Delafield's 

 hematoxylin enough acetic acid be added to give it a 

 red tinge. This will make it a more precise nuclear 

 stain. Cajal's Method for staining by diffusion or 

 propagation : Expose the brain of a rabbit and cut 

 sections of the cortex 2 mm. thick. Cover the sec- 

 tions on both sides with finely powdered methylene- 

 blue or with a saturated solution of the dye. Restore 

 the sections to the places from which they were cut and 

 close the skull for a half-hour. Remove the sections 

 and fix them in Bethe's ammonium molybdate for 2 

 hours; wash, harden for 3 or 5 hours in a mixture of 

 one part 1 % platinum chlorid, 40 parts formalin, and 

 60 parts water, and embed in paraffin. Treat the sec- 

 tions with alcohol containing 0.3^, platinum chlorid. 

 Cajal's Picroindigo-carmin, dissolve 0.25 gm. 

 indigo-carmine in 100 gm. saturated aqueous solution 

 of picric acid. Stain 'sections (previously stained in 

 carmine) for from 5 to IO minutes, wash in dilute 

 acetic acid, then in water, then in absolute alcohol. 

 Carazzi's Mixture. Dissolve 20 gm. of sublimate 

 in IOO c.c. of 70% alcohol, 15 c.c. of strong nitric 

 acid, and 5 c.c. of glacial acetic acid. Of this add 

 12 c.c. to 100 c.c. of IJ^ sodium chlorid solution. 

 Fix for from I to 6 hours, according to the size of the 

 object. Wash in iodin-alcohol (Zenker's fluid). 

 Carbol-kresyl Violet. See Morse's Method, Car- 

 min Blue, a cytoplasmic stain; used in acidulated al- 

 coholic solution (Janssen). Carnoy's Method for the 

 study of the structure of cytoplasm : Fix and stain with 

 methyl-green dissolved in 2^ or 3% aceticacid ; aftera 

 half-hour wash with acetic acid of the same percentage 

 and then substitute glycerol for the acid. Celli's 

 Method for the cultivation of protozoa: Cultivate the 

 ameba material in a petri-dish on Fttcus crispus prepared 

 with 5% of water (with or without bouillon), and 

 strongly alkalinized by adding 4 or 5 c.c. of saturated so- 

 lution of sodium carbonate to 10 c.c. of the dissolved 

 Irish-moss. When the cysts are ripe make cultures in 

 hanging-drops in filtered fucus and isolate the different 

 species of amebas. Celli-Guarnieri Method for 

 staining the parasite of malaria: Treat the fresh film 

 with a very dilute solution of methylene-blue in sterile 

 blood serum or ascitic fluid, lor double staining a 

 little eosin may be added. Chenzinsky's Stain 

 for blood : Concentrated aqueous solution of methylene- 

 blue and distilled water equal parts. To this is added 

 an equal quantity of 0.5% solution of eosin in 60% 

 alcohol. Stain blood-films 4 to 5 minutes. Red 

 blood corpuscles stain a rose-red, nuclei of leukocytes 

 blue, and malarial parasites blue. Chilesotti's Car- 

 min Stain for axis-cylinders: Mix I gm. sodium 

 acid amnio (Griibl'er) with y 2 grain uranium nitrate 

 and boil 12 hours with looc.c. water. Filter, and 

 before using add I r /r. hydrochloric acid. Sections 

 from Miiller's fluid will stnin in 5 to 10 minutes ; those 

 from formalin, freezing paraffin, and celloidin in 15 to 

 20 minutes; from Weigert's neuroglia fluid in •-£ to I 

 hour ; from Marchi in 2 to 4 hours. Then treat with 



water, alcohol, carbolxylol. Cholera Red Reaction. 

 See Nitrosoindol Reaction. Chromogen, a naph- 

 thalene compound prepared at the Hochst dye-works. 

 Used by Weigert for staining neuroglia. See Uei- 

 gert's Method. Ciaglinski's Method, the same as 

 Strobe's method, with fore-staining in safranin and 

 differentiating in water instead of alkali-alcohol. 

 Claudius' Method for bacteria : Stain in gentian or 

 methyl-violet, after Gram (see Gram's Method), dif- 

 ferentiate in a saturated aqueous solution of picric acid 

 diluted with an equal volume of water, decolor in 

 chloroform. Cleavage. Methods of study. 1. 

 Total equal or adequal cleavage. Use a slide with a 

 cell. Put a filament of sea-alga with a very little sea- 

 water in the cell and over it a cover-glass with a drop 

 of water containing fertilized eggs of echinoderms sus- 

 pended in it. Fix with osmic acid or liquid of Flem- 

 ming. 2. Total unequal or inequal cleavage. Fix 

 the eggs of the leech or the snail in Flemming's solu- 

 tion, stain in borax-carmin, and embed in paraffin. 

 3. Superficial cleavage. Suitable objects are the eggs 

 of the viviparous plant-louse. Eggs and embryos in 

 different stages of cleavage are obtained by teasing the 

 insect in physiologic salt solution; or the insect entire 

 may be killed in hot Mater, hardened in alcohol, and 

 embedded in paraffin. 4. Discoidal cleavage. Fix 

 the eggs of the cuttle-fish in picrosulfuric acid and dis- 

 sect off the germinal disc. 5. Influence of pressure. 

 Place a bristle of medium size on a slide and beside it 

 the fertilized egg of an echinoderm, in a drop of 

 water. Apply a cover-glass. All gradations of 

 pressure-effects can be observed between the bristles 

 and the far edge of the cover. For larger eggs — for 

 example, frogs' eggs — the procedure is as follows : Ce- 

 ment 2 strips of glass about 1. 4 mm. thick on the 

 edges of a slide. Place the eggs on the slide, cover 

 with another slide, and tie the two slides together. 

 The eggs may be fertilized before (Hertwig) or after 

 (Born) compression. Place some of the eggs in a ver- 

 tical, others in a horizontal position and observe the 

 deviating course of cleavage. Cf. Driesch's Method. 

 Coles' Method for staining the diphtheria bacillus: 

 Fix the films by heat or in absolute alcohol and ether 

 and stain in Neisser's methylene-blue (see A'eisser's 

 Method); wash, and treat with the 1:2:300 solu- 

 tion of iodin and potassium iodid; wash, and stain 

 in vesuvin. Time in each solution a half minute. 

 Conklin's Stain for the embryo chick : Mix equal 

 parts of Delafield's hematoxylin and distilled water 

 and add I drop of picric acid solution to each cubic 

 centimeter of the dilution. For use dilute with 4 vol- 

 umes of water and stain for from 10 to 20 minutes. 

 Conn's Method for preserving cultures of bacteria as 

 museum specimens: Inoculate 2% agar slants and 

 seal the tubes with paraffin and plaster of Paris. In a 

 few days the cultures cease growing and remain indefi- 

 nitely unaltered. Cook- Zimmerman Method for 

 histologic sections of the cochlea: Decalcify the 

 petrous bone of a kitten of about 2 weeks in 5 '7 nitric 

 acid; wash for 8 hours in water, for 3 hours in 

 alcohol, for 6 hours hi 50% alcohol. Cut thin slices, 

 parallel to the auditory nerve and the modiolus, and 

 treat them for 4 hours with "]o r / ( alcohol, with borax 

 carmin for 12 hours, and destain in acid alcohol. 

 Dehydrate, and embed in paraffin by the cedar oil pro- 

 cess. Attach the sections to the slide with albumen 

 fixative and treat them with xylol for 5 minutes (with- 

 out dissolving the paraffin) ; with absolute alcohol and 

 95$ alcohol each for 2 minutes, with < 5$ alcoholic 

 solution of picric acid for I minute, with 95% alcohol 

 and absolute alcohol each for 2 minutes, with xylol 

 for 5 minutes, and mount in balsam. Coming's 



