344 



METHODS AND FORMULAS 



DS 13.11 



be pushed across the lower slide, not pulled 

 across it. That is, one should lay one 

 chemically clean slide on the bench, place 

 a small drop of blood on it, and place a 

 second slide in contact with the first at an 

 angle of about 45° so that the blood is 

 drawn out by the capillary attraction into 

 the angle. The upper shde is now pushed 

 forward (Fig. 29, Chapter 7) leaving be- 

 hind it a thin smear of blood which is then 

 air dried. The blood film is fixed either by 

 placing it in chemically pure methanol for 

 from one-half to one minute, or by pouring 

 two or three doses of chemically pure 

 methanol over it. After removal from 

 methanol, the slide is dried, and the stock 

 staining solution A (DS 13.13 Kingsley 

 1935, below) is then poured on it and left 

 for a period of from five to eight minutes. 

 This time is, of course, too long for ordi- 

 nary rapid diagnostic work, but has the 



advantage that slides may be accumu- 

 lated in a long fine. Then one may con- 

 tinue the process on the first slide, when 

 one has spent five minutes in preparing 

 the series. After tlie required time has 

 elapsed, the stain is washed off with a jet 

 of distilled water directed on it from a 

 wash bottle in such a manner as to float 

 off the scum which has gathered on the 

 surface as well as to rinse off the su- 

 perfluous stain. The film may then either 

 be air dried for examination, or it may 

 be mounted in a neutral mounting me- 

 dium (see Chapter 26) after dehydration 

 through direct drying. Provided the stain- 

 ing solution has been accurately made and 

 pure reagents used, a perfect stain wiU in 

 every instance result from this treatment. 

 The methods given for paraffin sections 

 are just as easy to follow and need not be 

 elaborated here. 



13.11 METHYLENE BLUE-EOSINATES 



13.11 Assmann 1906a test. 1928 Schmorl Schmorl 1928, 241 



REAGENTS REQUIRED: ^4. DS 13.11 May-Grunwald 1902 (working sol.); B. water 100, 



DS 11.44 Unna 1892 

 method: [dried smear] -^> A, 1 ml. poured on slide lying in petri dish, 3 mins. 

 15 ml. poured into dish around slide, 3-4 mins. -^ wash — > dry 



B. 



13.11 Assmann 1906b test. 1928 Schmorl Schmorl 1928, 246 



REAGENTS REQUIRED: A. DS 13.11 May-Grunwald 1902 (working sol.); B. 0.001% acetic 



acid 

 method: [sections of F 7000.0000 Miiller 1859 or F 3700.0010 Zenker 1894 fixed ma- 

 terial] — » A, several hrs. -^ B, till color changes to clear eosin -^ wash — > balsam, via 

 usual reagents 



13.11 Chenzinsky 1894 23632, 11 :269 



formula: sat. sol. (circ. 4.5%) methylene blue 40, 0.5% eosin Y in 70% ale. 20, water 20 

 method: [fresh smear] — * stain, 5 mins. -^ water, rinse —>-^ dry 



13.11 Ellerman 1919 23632, 36:56 



reagents required: A. water 100, 40% formaldehyde 5, eosin Y 1; B. DS 13.11 May- 



Griinwald 1902 (working sol.) 50, water 50 

 method: [5 M paraffin sections of F 3700.1000 Ellerman 1919 fixed material] — > water — » 

 A, 15 mins. — > wash, 2-4 mins. 45°C. — > B, 30 mins. -^ wash, 5-10 mins. — > blot — > 

 abs. ale. till differentiated — ^ M 32.1 mountant via usual reagents 



13.11 Held (1900) see DS 22.21 Held (1900) 



13.11 Jenner 1899 11995, 6:370 



preparation of dry eosinate: Mix equal parts 1.25% eosin Y and 1% methylene 



blue. Leave 24 hours. Filter. Wash and dry filtrate. 

 WORKING solution: dry powder 0.5, methanol 100 

 method: [fresh smear] — » stain, 3 mins. — > water, rinse — > dry 

 NOTE : The most usual employment of this formula is as a fixative before such methods 



as DS 13.13 Slider and Downey (1929). 



13.11 Jenner test. 1905 Lee Lee 1905, 385 



formula: 0.5% methylene blue 50, 0.5% eosin Y in methanol 62.5 

 method: as Jenner 1899 



