VAGINAL SMEARS 



368 



VAN DEN BERGH TEST 



birefringent; and in the dark field the 

 surface was brilliantly illuminated. 

 They did not take colors dissolved 

 weakly in the mounting fluid. When 

 Janus Green B was offered in weak di- 

 lution, coccoid mitochondria were ob- 

 served between the vacuoloids. Fine 

 red-stained granules were found be- 

 tween the vacuoloids in foam cells of 

 frozen sections which had been stained 

 with Sudan IV. Vacuoloids seem to be 

 composed largely of water, but should 

 not be regarded as simple capsules. 

 They should certainly not be confused 

 with myelin figures. When foreign 

 particles are taken into the cell they 

 lodge between, never within, the vacuo- 

 loids, and this is true for vital dye accu- 

 mulations. The possibility of differen- 

 tiation in vacuoloids is admitted. 



In material fixed in Regaud's fluid, 

 mordanted in potassium bichromate, 

 cut into frozen sections and well stained 

 with Heidenhain's iron hematoxylin, 

 some of them appear as dense blue- 

 black balls. Some of them appear as 

 large dense darkbrown spheroids or 

 ovals within pneumonocytes which 

 have been in contact wath blood and 

 have been fixed with pure formalin or 

 other aldehyde (Sjostrand and Sjos- 

 trand, Zeitsch. f. mikros.-anat. Forsch., 

 1938, 44, 370-411). These workers find 

 that the colored material has properties 

 of hemin. The same coloration of 

 vacuoloids may occur in mice suffering 

 from a virus infection after fixation 

 with Bouin's fluid by immersion of the 

 skinned intact thorax (which see). 

 Some of them, in apparently normal 

 pneumonocytes and foam cells appear 

 black or gray-black after exposure to 

 osmic acid fixation intrabronchially 

 after Aquax (which see) embedding and 

 mounting in levulose (which see). In 

 apparently degenerate pneumonocytes, 

 all of the vacuoloids are impregnated 

 with osmium. 



The presence of vacuoloids brings 

 about a lacelike picture in the mito- 

 chondria of pneumonocytes, for mito- 

 chondria are not admitted to the in- 

 teriors of the vacuoloids but occupy the 

 cytoplasm between them (Macklin, 

 C. C., Biol. Bull., 1949, 96, 173-178). 

 With Nile Blue Sulphate (Lorrain 

 Smith, which see) the vacuoloids in 

 foam cells which are not stained dark 

 after osmic acid fixation are some- 

 times colored light red as seen in 

 Aquax sections mounted in levulose. 



Macklin spoke in the following 

 words at the 1947 International Cyto- 

 logical Congress: "The function of the 

 foam cell is obscure, and may be con- 

 cerned with the conditioning of the 



alveolar wall; as for instance in the 

 maintenance of a proper surface ten- 

 sion. However, the finding of these 

 cells in augmented quantities under 

 pathological conditions in man and 

 rat suggests that they may be impli- 

 cated in the local body reactions to 

 chronic infection. Amoeboidism seems 

 to be a property. ... It looks as if the 

 antecedent entodermal cell has a dual 

 differential potency, directing it to the 

 left, so to speak, where it may become 

 an irreversible pure foam cell, or to the 

 right, where it may end up as an irre- 

 versible pure dust cell. The in-be- 

 tween stages, presumably very useful, 

 may be termed "bireactives." (Mack- 

 lin, C. C., Proc. 6th Internat. Cong, 

 exp. Cytol., Stockholm, 1947; published 

 1949, 383-387). 



Vaginal Smears, see Papanicolaou Tech- 

 niques. 



Valves. Aortic, staining of elastic tissue 

 in (Wilens, S. L., Arch. Path., 1940, 

 29, 200-211). X-ray demonstration of 

 valves of veins (Edwards, E. A., Anat. 

 Rec, 1936, 64, 369-385). 



Vanadium, see Atomic Weights. 



Van den Bergh Test for bilirubin as de- 

 scribed by Wintrobe, M. M., Clinical 

 hematology. Philadelphia: Lea & 

 Febiger, 1942, 703 pp. abbreviated: 



1. Qualitative: 



(a) Add 1.5 cc. cone, hydrochloric 

 acid C.P. to 30 or 40 cc. aq. dest. -f 

 0.1 gm. sulphanilic acid which keeps 

 well. 



(b) Dissolve 0.5 gm. sodium nitrite 

 C.P. in 100 cc. aq. dest. making up fresh 

 every 3-4 weeks. 



Make diazo reagent by mixing 5 cc. 

 of (a) with 0.15 cc. of (b) freshly each 

 day. 



Mix 0.25 cc. reagent with 0.2 cc. clear 

 plasma or serum (2 cm. column in hema- 

 tocrit). Immediate purplish color at- 

 taining maximum in 30 sec. is direct 

 reaction. Color appearing at once but 

 reaching maximum later is biphasic re- 

 action. If no color in 1 min. but on 

 addition of 5 cc. alcohol reddish violet 

 color appears reaction is indirect. 



2. Quantitative. 



(a) Stir and shake 80-90 gms. am- 

 monium sulphate C.P. in 100 cc. aq. 

 dest. until saturated and filter. 



(b) Make standard color by dissolving 

 3.92 gm. cobalt sulphate (7H2O) in 100 

 cc. aq. dest. over night. 



Mix 0.5 cc. diazo reagent with 1 cc. 

 serum or plasma in centrifuge tube. 

 After standing few minutes add 2.5 cc. 

 95% ethyl alcohol and 1 cc. of (a). Mix 

 and centrifuge. In positive reaction 

 uppermost layer is reddish violet alco- 

 holic extract of diazotized bilirubin, 



