784 SUMMARY OF CURRENT RESEARCHES RELATING TO 



Absence of Altmann's Granules in Cancer Cells.* — H. Beckton 

 adopted the following procedure in an investigation as to the absence of 

 Altmann's granules from cells of malignant new growths : — 1. Take 

 small pieces of tissue as soon as possible after an operation. 2. Place 

 in formol-Miiller (1 in 49) at room temperature for a week, renewing 

 the solution on the second and fourth days, and then in Muller's fluid 

 for another week. Usually for very small pieces, 1 mm. thick, 3 days 

 are sufficient. 3. Wash overnight in running water ; a few hours 

 is sufficient time for very small pieces. 4. Alcohol in increasing 

 concentration ; cedar-wood oil or xylol ; paraffin, 75° C. 5. Cut sections 

 not thicker than 5/x. and mount on slides. 6. Xylol ; absolute alcohol ; 

 rinse in water. 7. Stain in anilin-acid-fuchsin. 8. Differentiate with 

 picric-acid alcohol or with aqueous solution of sodium bicarbonate 

 (1 in 10,000). 9. Absolute alcohol ; xylol ; Canada balsam. 



The stain is prepared thus :— To 100 c.o. of filtered cold saturated 

 watery solution of anilin add 20 grams of acid-fuchsin ; shake well and 

 filter. The picric acid alcohol is made by adding 7 volumes of 20 p.c. 

 alcobol to 1 volume of saturated picric acid solution in absolute alcohol. 



The process of staining may be carried out at room temperature, the 

 slide being placed vertically in the staining fluid for, say, half-an-hour. 

 This method is free from any objection which may be urged against 

 the use of a hot staining fluid. The following method, however, 

 gives the same results and is most expeditious and convenient. Cover 

 the section with staining fluid, and this in turn with a watch-glass, 

 and beat for 2 minutes to about 60° C, using either a thermostat or 

 a strip of copper heated at one end and tested from point to point by 

 means of a piece of paraffin wax. Next pour off the stain, blot, and 

 differentiate ; a point is soon reached at which but little further colour 

 comes away. As a rule, at ordinary temperatures differentiation is 

 complete with picric-acid alcohol in about 2 minutes, and with sodium 

 bicarbonate in about J minute. 



It may be noted here that absolute alcohol does not decolorise a 

 section, but dilute alcohol discharges the colour rapidly, while plain 

 water removes it but slowly, and indeed differentiates sufficiently to 

 enable one to detect the presence of granules. Again, while in 

 moderately dilute mineral acids the colour remains in a section for a 

 long period, moderately dilute alkalis rapidly discharge it, and very 

 dilute alkalis may be used as differentiating fluids. 



After fixation with formalin alone, as well as after formol-Miiller 

 solution, granules can be demonstrated in the essential cells of pancreas, 

 submaxillary gland, liver, kidney, columnar epithelium of the alimentary 

 tract, etc. ; but this is not the case with lymphocytes, plasma cells, 

 endothelial cells, fibroblasts, fat-cells, etc., which show grannies only 

 after fixation with formol-Miiller. It thus appears that the granules 

 by this method may be appropriately referred to as belonging to a 

 " gland cell group " and a " connective-tissue cell group " respectively. 



The conclusion drawn from the results of this investigation is, that 

 in malignant new growths granules tend to disappear, or to be absent 

 altogether, from the particular type of cell involved. 



* Brit. Med. Joum.,1909, ii. pp. 859-61 (3 figs.). 



