UROBILIN 



258 



VAN GIESON'S CONNECTIVE 



urobilinogen not converted into uro- 

 bilin bj' adding 1-2 drops of Lugol's 

 Iodine. Then H,dd 10 cc. Schleslnger's 

 Reagent, filter let stand 1-2 lirs. Uro- 

 bilin is indicated by green fluorescence 

 when examined against dark back- 

 ground in bright light. 



2. Make dilutions of urine by adding 

 1 cc. to 10, 20, 30, 40 etc. cc. of aq. dest. 

 To 10 cc. of each dilution in test tubes 

 add 1 cc. Ehrlick's Aldehyde Reagent. 

 Urobilinogen is indicated by pink color 

 within 5 min. seen by looking down 

 through mouths of tubes. 



Vaccinia, Cytoplasmic inclusions in, see 

 Cowdry, E. V., J. Exper. Med., 1922, 

 36, 667-684. Summary of methods used 

 in the investigation of elementary 

 bodies of vaccine virus (Smadel, J. E. 

 and Hoagland, C. L., Rev. Bact., 1942, 

 6, 79-110. 



Vaccinia, see Guarnieri Bodies. 



Vaginal Smears. On the basis of large 

 experience Papanicolaou G. N., J. Lab. 

 & Clin. Med., 1940-41, 26, 1200-1205 

 has described techniques in detail. 



1. Fix immediately (before drying) 

 in equal parts 95% alcohol and ether 

 1-2 min. Rinse in 70%, 50% alcohol 

 and in aq. dest. Ehrlich's hematoxylin 

 (or other hematoxylin), 1-2 min. Rinse 

 in aq. dest. Rinse few times in 1% 

 hydrochloric acid (may be omitted). 

 Running water, 5 min., or aq. dest. 100 

 cc. + 3 drops cone. aq. lithium car- 

 bonate, 1 min. Do not leave slides too 

 long in running water. Rinse in aq. 

 dest. and stain for 2 min. in any of 6 

 combinations of stains recommended. 

 One of these is made up of National 

 Aniline and Chemical Co. dyes in 0.5% 

 aq. sol. as follows: Light green S. F. 

 yellowish, 12 cc. ; orange G, 24 cc, acid 

 fuchsin, 20 cc, eosin yellowish, 40 cc. -f 

 phosphomolybdic acid (Merck) 0.45 

 gm. Rinse in water. Rinse in dioxan 

 10-15 times until smears are clear. 

 Pass through absolute alcohol to xylol. 

 Mount in clarite, balsam or dammar. 

 See a newer procedure by Papanicolau, 

 G. N., Science, 1942, 95_, 438-439. _ 



2. A shorter method is, after similar 

 fixation of smears brought down to aq. 

 dest., to stain them 2-3 min. in anilin 

 blue, water soluble, 16 cc. ; acid fuchsin, 

 23 cc. ; orange G, 17 cc. ; eosin yellowish, 

 44 cc, (all of 5% aq. solutions) + phos- 

 phomolybdic acid 0.2 gm., and phos- 

 photungstic acid, 0.2 gm. Rinse in 

 water. Rinse in dioxan until clear, 

 then through absolute alcohol and 

 xylol to clarite. Nuclei, red; erythro- 

 cytes, orange ; cornified cells, red, pink 

 or orange; basophile cells, green or blue. 

 Nuclei not as dark and cell outlines as 

 sharp as after hematoxylin, but corni- 



fied cells are more prominent and basal 

 cells more transparent. 



3. See detailed method advised by 

 George Svihla for the rat (Hartman, 

 C. G., Yale J. Biol. & Med., 1944, 17, 

 99-112). 



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. + 

 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, 

 next layer is flocculated protein and 

 residue is ammonium sulphate. Com- 

 pare supernatant fluid with the stand- 

 ard (b) in colorimeter. Then: 



mm. standard 



; X 4 X 0.5 



mm. unknown 



= mg. bilirubin per 100 cc. 



Van Gehuchten's mixture, see Carnoy's 



Fluid. 

 Van Gieson's Connective Tissue Stain. 



Paraffin sections of Zenker fixed mate- 

 rial are stained with Harris' hema- 

 toxylin. Rinse in water. Stain in 1% 

 aq. acid fuchsin 7.5 cc and sat. aq. 

 picric acid 50 cc, 10 min. Wash in 



