450 METHODS AND FORMULAS DS 22.4-DS 22,6 



note: The reflux boiling should continued until a xylene extract of a sample shows 

 strong red fluorescence. The chemical validity of this separation has been sharply 

 queried by Kaufmann and Lehmann 1926 (22575, 261:623). For a discussion of the 

 arguments for and against see Gatenby and Cowdry 1937, 280 and Cowdry 1943, 136. 

 The fact remains that fatty cell inclusions are differentiated in red and blue, on a 

 reproducible basis. 



22.4 Smith and Rettie 1924 11431, 27:115 



BEAGENTS kequired: A. ADS 12.1 Smith and Rettie 1924; B. water 99.5, acetic acid 



0.5, hematoxylin 1; C. ADS 21.1 Smith and Rettie 1924 

 method: [frozen sections of formaldehyde-fixed material] -^ A, 1-2 days, 37°C. — > wash 

 -^ B, 6-18 hrs. -^ C, till differentiated-^ wash— > M 12.1 mountant 



22.4 Vrtis 1931 23632, 47 :443 



REAGENTS REQUIRED: A. 4% formaldehyde; B. sat. sol. Sudan III in 70% ale. 

 method: [pieces of skin] —> A, overnight -^ wash ^50% ale, J^ to 1 hr. — > B, }i-l hr. 



— > 50% ale, till differentiated — > glycerol or M/10 mountant. 

 RECOMMENDED FOR: fat glauds in wholemounts of rodent skin. 



22.4 Wilson 1950 4349, 31:216 



reagents required: A. DS 22.4 Lillie and Ashburn 1943; B. DS 11.124 Gomori 1941; 



C. 1% acetic acid; D. 0.5% light green 

 method: [5 II frozen sections, mounted on slide] -^ A, 10 mins. -^ thorough wash — > B, 



4 mins. —* tap water, till blue -^ C, rinse -^ D, quick dip — > C, rinse — > M 12.1 Kaiser 



1880 



22.5 Plastics 



Plastids are commonly demonstrated in sections of plant tissue by a hematoxylin staining 

 technique, or by any other technique which is customarily employed in the histological 

 examination of vegetable structures. The two methods given below are those which dis- 

 tinguish (if distinction is possible) between proplastids and mitochondria. The method of 

 Milovidov 1928 is merely a modification of the standard Altmann acid fuchsin-aurantia 

 mitochondrial staining technique, while the technique of Nemefi 1906 relies on prior mordant- 

 ing in tannin before the use of gentian violet. 



22.5 Milovidov 1928 1823, 24:9 



reagents required: A. DS 22.2 Altmann 1890 (A. sol.); B. 5% aurantia in 95% ale; 

 C. 2% tannin; D. 1% methyl green 



method: [sections of F 7000.1000 Regaud 1910 (or any F 6700.1000 fixative) fixed 

 material] —> A, poured on sUde and warmed till steaming, 1 min. — > B, till mito- 

 chondria alone remain red — » 70% ale, rinse -^ C, 20 mins. -^ D, 5-10 mins. -^95% 

 ale, till starch grains sharply differentiated -^ balsam, via usual reagents 



result: mitochondria and proplastids, red; starch, green. 



22.5 Nemec 1906 2626,24:528 



reagents required: A. 2% tannin; B. 1.5% antimony potassium tartrate; C. 1% 



gentian violet 

 method: [paraffin sections of F 5000.0015 Nemec 1899 fixed material]-^ waters A, 



1 hr. -^ water, wash -^ B, 5-15 mins. — > water, wash — > C, ^i to 2 hrs. — ^ water, 



wash -^95% ale, till starch granules well differentiated -^ balsam, via usual reagents 

 note: Schneider 1922, 351 recommends fixation in F 1600.0010 Flemming 1882, with 



subsequent bleaching in hydrogen peroxide. 



22.5 Schneider 1922 see DS 22.5 Nemec 1906 (note) 



22.6 Staech, Glycogen, and Amyloid Granules 



Starch and glycogen are both most commonly stained by the application of iodine, 

 which turns the former blue and the latter brown. These iodine techniques, however, do 

 not leave permanent stains, though endeavor has been made by Reilhes 1941 to provide 

 a degree of permanence in a resin mount. These preparations do not last, at their best, 

 more than a few months. The original method for the differential staining of glycogen 



