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contents of the pipet into a sterile test-tube. Prepare 

 another dilution in which the blood is drawn to the 

 0.5, 0.3, or O.I mark. Centrifugate the test-tubes. A 

 drop of the serum of each is then examined by the 

 hanging-drop method. As 100- volume parts of blood 

 contain 67.762 parts of serum, the volume of serum is 

 practically equal to two-thirds that of the blood. 

 When the blood is drawn to the mark o. I , the dilution 

 is I : 150 ; when to the mark 0.5, I : 30, and when to 

 the mark 1.0, 1 : 15. Pfeiffer's Method for bacteria 

 in tissue sections : Harden in alcohol and stain for a 

 half hour in carbol-fuchsin solution and distilled water 

 I : 20. Wash in distilled water acidified with acetic 

 acid. La Phenicienne. See Bismarck-brown under 

 Staining Reagents (Illus. Diet.). Pick's Method 

 of preserving tissues: I. Place specimen for one or 

 two days in Pick's formalin salt solution, consisting of 

 distilled water, ico parts; sal carolin factit, 5 parts; 

 formalin, 6 parts. Sal carolin factit is composed of 

 potassium sulfate, 2 parts; sodium chlorid, 15 parts; 

 sodium bicarbonate, 33 parts; sodium sulfate, 44 

 parts. 2. Place the specimen for 24 hours in 80% to 

 90% alcohol. 3. Place the specimen promptly in 

 Kaiserling's solution: acetate of potash, 5 parts; gly- 

 cerin, 10 parts; distilled water, 1 00 parts. This solu- 

 tion may be varied in strength even up to the concen- 

 trated form used by Melnikow-Raswedenkow, which is 

 acetate of potash, 30 parts; glycerin, 60 parts; dis- 

 tilled water, 100 parts. Pick-Jacobson Method for 

 bacteria: Stain from 4 to 10 seconds in a mixture of 

 carbol-fuchsin 15 drops, concentrated alcoholic solution 

 of methylene-blue 8 drops, distilled water 20 c.c. 

 Bacteria dark blue, nuclei light blue, protoplasm and 

 mucin red. Picric-acetic Acid for fixing tissues: 

 Saturated solution of picric acid 100 c.c. and glacial 

 acetic acid I or 2 c.c. Place the tissue in this for from 

 6 to 12 hours, then in 70% alcohol for one day, and 

 then in 80% alcohol, renewed as often as it becomes 

 yellow. Picric Alcohol, a saturated solution of pic- 

 ric acid in 50% alcohol. Picronigrosin, a solution 

 of 1 gm. of picric acid in loo c.c. of distilled water 

 with the addition of 1 gm. of nigrosin. Pierce's 

 Method for sealing cultures of organisms that grow on 

 potatoes : Thrust loose cotton to the bottom of the tube 

 to the depth of an inch and pour in distilled water to 

 the depth of a half inch. Drop a potato plug on the 

 cotton and close the tube with a cotton plug, in the 

 usual way. Steam for an hour. Inoculate, and when 

 the cultures are satisfactory trim the cotton plug, flame 

 it, and then push it into the tube for a distance of one- 

 eighth inch. Pour a little melted paraffin on the cotton, 

 and when this has hardened fill the space above the 

 cotton with paraffin. Piorkowski's Medium. I. 

 For the cultivation of bacilli belonging to the colon 

 group: Add 0.5% of peptone and 3.3% of gelatin 

 to urine that has acquired the alkaline reaction, and 

 cook for one hour in a water-bath. Filter, and ster- 

 ilize in test-tubes for 15 minutes in a steam bath, 

 and again for 10 minutes on the following day. 2. For 

 differential staining of diphtheria bacilli : Make dry 

 cover-glass preparations of a culture on Isomer's blood - 

 serum, at a temperature of 37 C. for 20 hours and 

 stain 30 seconds in methylene-blue; decolor in 3% 

 hydrochloric acid for 5 seconds and counterstain in 1 ', 

 aqueous solution of eosin for 5 seconds. Pitfield's 

 Method for staining spores : Fix the film in flame 

 and stain in boiling carbol-fuchsin or in Ehrlich anilin 

 gentian-violet ; wash, and decolor with a drop of a so- 

 lution of ammonium persulfate 5 gm., in 50 c.c. of 

 95% alcohol and 10 c.c. of water ; after a half minute 

 wash and counterstain. Plato's Method for staining 

 gonococci in living leukocytes : Add I c.c. of a cold 



saturated aqueous solution of neutral red to 100 c.c. 

 of physiologic salt solution. Mix a small drop of the 

 fresh gonorrheal pus with the stain and examine in a 

 hanging drop. Plaut's Method for the bacilli of 

 diphtheria : Stain in dilute carbol-fuchsin or in a 

 mixture of 5 parts of concentrated alcoholic gentian- 

 violet solution and 95 parts of anilin water ; decolor in 

 alcohol, or better in anilin. Plehn's Method. 1. 

 For the study of the living malarial parasite : Place a 

 drop of fluid paraffin on a slide and a drop on a cover- 

 glass ; take up the drop of blood on the latter and so 

 place it on the slide that the blood is between the 

 drops of paraffin. Examine on a warm stage. The 

 addition of a drop of methylene-blue will stain the 

 living organisms. 2. For malarial films fixed in abso- 

 lute alcohol : Stain for 5 minutes in a mixture of con- 

 centrated aqueous solution of methylene-blue, 60 c.c, 

 0.5% solution of eosin in 75% alcohol, 20 c.c, dis- 

 tilled water, 20 c.c, and 20^) potash lye, 12 drops. 

 Plehn's Stain for blood. (This is a modification of 

 Chenzinsky's stain.) Concentrated aqueous solution 

 of methylene-blue, distilled water, equal parts. To 

 this add one-half the equal volume of ao.5^ solution 

 of eosin in 60% alcohol. .Filter before use. Fix 

 blood-films in absolute alcohol for 7 to 10 minutes. 

 Stain from a few minutes to 24 hours. Red blood- 

 corpuscles and eosinophil granules stain a rose-red. 

 The nuclei of leukocytes stain a dark -blue, and mala- 

 rial parasites a light-blue. Polychrome Methylene- 

 blue, a reddish -violet dye sometimes present as an im- 

 purity in commercial methylene-blue, or that develops in 

 old, ripened or alkaline solutions of methylene-blue. It 

 is used for staining cell granules. See L'nna's Method. 

 Pommer's Method for the study of the deposition of 

 calcareous substances in bone and for the detection of 

 nonnucleated areas : Treat the bone with Miiller's 

 fluid until it can be cut with a razor. In the sections 

 the previously calcareous areas are recognized by their 

 homogeneous appearance, the noncalcified portions 

 by their fibrillar structure. It is of advantage to stain 

 these sections with carmin. For the staining of bone 

 that has been decalcified by an acid 0.04^ solution of 

 dahlia, or O. I Jj solution of safranin, or 0.3^ solution 

 of methyl green may be used. From 12 to 18 hours 

 are necessary for sections. The areas that previous to 

 decalcification were calcareous will be intensely col- 

 ored, the areas previously noncalcified will be color- 

 less. Primrose Soluble, a phthalein dye resembling 

 eosin. It is not wholly identical in properties, but va- 

 ries according to the mode of manufacture. Prince's 

 Stain : Prepare a mixture of 2 parts of 2 c / ( solution 

 of eosin, one part of saturated solution of acid fuch- 

 sin, and 24 parts of saturated solution of toluidin blue; 

 agitate and decant. In the fresh solution films stain in 

 a few seconds ; in a few minutes after it is several 

 weeks old. Progressive staining of Heidenhain, a 

 method in which the pigment used is one that will 

 stain some tissue elements or one structural part of 

 a cell more rapidly than others and in which the 

 process of staining is arrested before the remaining ele- 

 ments become colored. It is the same as the "di- 

 rect" method of Flemming. Cf. Regressive Staining. 

 Pyronin-methyl Green, a stain consisting of I % so- 

 lutions of pyronin and methyl green in distilled water 

 made separately and mixed ; 4 parts of the former to 1 

 of the latter. Quincke's Method for obtaining the 

 ammonium sulfate reaction in ferruginous tissues: The 

 author advises instead of fresh solution of ammonium 

 sulfate that which has become yellow with age. Con- 

 centration of the solution or the addition of a little 

 ammonia sometimes accelerates the initiation of the 

 reaction. Cf. the methods of Hall and Zalewski. 



