STAINS 



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STAINS 



for the cultivation of comma bacilli : Dilute the in- 

 testinal contents with an equal volume of alkaline 

 bouillon and expose to air for 12 hours. The bacilli, 

 owing to their necessity for oxygen, develop chiefly on 

 the surface, v. Schrotter's Method of staining 

 the medullary sheath : The sections, which are best 

 hardened in Muller's fluid, are placed from 15 to 20 

 minutes in a freshly prepared cold solution of gallein 

 (Griibler), which is prepared by boiling with well- 

 water. Then differentiate in a \f solution of soda 

 or weak sodium hydroxid solution, then for a moment 

 in a light violet permanganate solution. Wash with 

 water, absolute alcohol, carbol-xylol. The medullary 

 substance will have a violet appearance, likewise the 

 red-blood-corpuscles; the gray substance and connec- 

 tive tissue will remain unchanged. Schultze's (O.) 

 Method for the preparation and preservation of trans- 

 parent embryos : Harden the embryo in alcohol and 

 transfer directly to a 3 % or 5 % aqueous solution of 

 caustic potash. In about a week the embryo be- 

 comes transparent and is then preserved in a mixture 

 of glycerol 30 parts, formalin 2 parts, water loo parts. 

 Treatment with potash solution alone will make the 

 tissues transparent and isolate the bones, but the 

 preparation cannot be preserved. Schultze's Method 

 for smooth muscle : Treat for 24 hours with 10% 

 nitric acid, wash, and treat for a week (in the dark) 

 with a mixture of equal volumes of 0.05% osmic acid 

 and 0.2% acetic acid; tease and mount in glycerol. 

 Shaffer-Bouma Method for cartilage: Stain for from 

 30 to 60 minutes in 0.05% aqueous solution of safra- 

 nin ; wash in water ; treat for 2 or 3 hours with o.ifo 

 solution of sublimate ; transfer to alcohol, dry with 

 filter-paper, and clear for a long time in clove or ber- 

 gamot oil. This method is also applicable to bone 

 that has been decalcified in nitric acid. See also 

 Zachariade 1 s Method. Siemerling's Method for 

 histologic preparations of the brain : Harden in a mix- 

 ture of Miiller's fluid 100 parts and formalin 2 parts. 

 Treat the sections with 0.55% solution of chromic acid 

 and stain after Weigert's method. Silk-thread Test: 

 Sterilize pieces of silk thread, I cm. long, and dip 

 them into a suspension in sterilized water of the bac- 

 teria to be tested ; after a few minutes transfer the 

 threads to a sterilized petri dish, and when dry dip 

 them into a solution of the disinfectant to be tested. 

 Remove them one by one, at intervals of 5» IO > 1 5> 

 30, and 60 minutes, and transfer them to tubes of 

 nutrient bouillon. Sjobring's Method for fixing 

 tissues with formaldehyd : It is important to use the 

 formol of Meister, Lucius u. Briining. Treat mam- 

 malian tissue for 2 days with formol diluted with 4 

 volumes of water and then transfer into 95 °/ c alcohol, 

 in which the object should remain for 2 days. If the 

 tissue contains much water, the hardening should be 

 begun in dilute alcohol. Formol is not advised for 

 fixing kinetic nuclei and is said to lessen the capacity 

 of nerve-tissue for taking stains. Smith's (Grieg) 

 Method for double-staining spores and bacilli : Dis- 

 tribute the bacteria in normal salt solution in a test- 

 tube, add an equal volume of carbol-fuchsin, and place 

 in boiling water for 15 minutes. Spread a loopful on 

 a cover-glass, dry, and fix in flame ; decolor in alcohol 

 containing 1.5% hydrochloric acid, wash, and stain in 

 methylene-blue. Smith's (S.) Method for staining 

 sections before dissolving out the paraffin : I'ut the 

 stain in a shallow, open dish. Float the ribbons 

 of sections on the stain. Stand the dish in a warm 

 place until the sections are flat, then cover it to prevent 

 evaporation ; after 24 hours pour off the stain, treat 

 with other necessary reagents in the same manner, 

 mount on the slide, and then clear and remove the 



paraffin with xylol or other clearing medium. In this 

 way thinner sections can be handled and attaching to 

 the slide is unnecessary. Sodium Dioxid for bleach- 

 ing tissue: Prepare a \o c / solution of tartaric or 

 acetic acid ; by means of a pipet introduce a little 

 sodium dioxid (Na 2 2 ) at the bottom of the container 

 and then cautiously pour on to the surface of the liquid 

 70% alcohol. Suspend the objects to be bleached 

 (previously saturated with alcohol) in the supernatant 

 alcohol. Solger's Method for centrosomes : In the 

 dermal pigment cells of the frontal and ethomoidal 

 region of the pike the centrosome may be seen without 

 staining. Fix in the liquid of Flemming. Heiden- 

 hain's iron-hematoxylin method will stain the centro- 

 some. Souza's Medium for fixing and hardening 

 tissues. See Pyridin. It also dehydrates and clears. 

 Sperm Crystals. To obtain these crystals when they 

 are present in pus, treat the exudate with salt solution 

 for 2 days and then add neutral ammonium phosphate. 

 Decant the supernatant liquid and examine the sedi- 

 ment. Cf. Bottcher's Method. Stabilit, a sort of 

 vulcanite manufactured for electric insulation and 

 recommended by Jelinek as blocks for mounting cel- 

 loidin objects. Steinschneider-Galewski Method 

 for gonococci : Stain for a half-hour in anilin-gentian- 

 violet, rinse, and treat for 5 minutes with solution 

 of potassium iodid, bleach in alcohol, rinse, dry, 

 and stain in alkaline methylene-blue. Stepanow's 

 Method. I. For embedding in celloidin : Dissolve 

 1.5 gm. of celloidin in 5 c.c. of clove oil, 20 c.c. of 

 ether, and I c.c. of absolute alcohol, added drop by 

 drop. Infiltrate in a stoppered bottle for from 1 to 6 

 hours, according to the size of the object ; uncork the 

 bottle and let the solution evaporate from 4 to 6 hours, 

 protected by a bell-jar; turn object and mass into a 

 silk-paper filter freely suspended in a warm place. 

 After from 4 to 6 hours cut out the object. Treat for 

 from 2 to 6 hours with vapor of chloroform and cut 

 with the knife dry. For dry sections preserve in 

 cedar oil, for wet sections in 85$ alcohol. The chief 

 advantages of this method are the transparency of the 

 mass and the rapidity of the process. 2. For the 

 bacilli of rhinoscleronia in tissue sections : Stain for 

 from 15 minutes to one hour in carbol -gentian violet 

 or for 24 hours in Loffler's methylene-blue, and ex- 

 tract briefly in alcohol containing o. i c / ( acetic acid. 

 Stephen's Method for fiagella : This is the same as 

 the method of Van Ermenghem, except that a 2<fo 

 solution of largin is used instead of silver nitrate. 

 Stephens- Christopher Method for preparing films 

 of malarial blood : Prick the finger with a triangular 

 surgical needle, touch the exuding drop with a slide, 

 and with the shaft of the needle spread the blood in a 

 broad, even streak, allowing time for the blood to run 

 along the edge of the needle by capillarity. Stieda's 

 Method for the detection of iron in tissue elements: 

 Stain sections of material hardened in alcohol or 

 formalin for several hours in lithium carmin ; wash 

 in water ; treat for from 4 to 6 hours with 2', potas- 

 sium ferrocyanid ; treat from 6 to 12 hours with \ c ' f 

 hydrochloric acid ; wash in water ; dehydrate, clear, 

 and mount in balsam. Strasburger's Method for 

 facilitating the sedimentation of urine or other secre- 

 tions containing bacteria : Mix one part of the secre- 

 tion with two parts of 95% alcohol. '1 he alcohol 

 causes the bacteria to sink by diminishing the specific 

 gravity of the liquid. The same method may be ap- 

 plied in the examination of feces for tubercle bacilli. 

 Strobe's Method for axis-cylinders : Harden in 

 Muller's fluid. Stain the sections for from 30 to 60 

 minutes in saturated aqueous solution of anilin-blue, 

 wash and transfer into filtered alkali-alcohol (caustic 



