PAPANICOLAOU TECHNIQUES 



256 



PAPANICOLAOU TECHNIQUES 



Gastro- Intestinal Tract. 



Esophageal aspirates are obtained by 

 means of a rubber suction tube passed 

 through the esophagoscope. Due to 

 the scantiness of material usually ob- 

 tained, the tube should be rinsed with 

 saline and this rinse added to the as- 

 pirate. The entire specimen is then 

 mixed immediately with 95% alcohol. 



Gastric specimens should be obtained 

 after at least eight hours of fasting. 

 Because of the action of free acid and 

 digestive juices in the stomach, best 

 results are obtained if the residual 

 gastric contents are first aspirated and 

 discarded. Freshly exfoliated material 

 can then be obtained by either of the 

 following methods. (1) Using a Reh- 

 fuss tube with a bucket tip, the stomach 

 is washed repeatedly with Ringer's 

 solution by aspirating and re-injecting 

 the same solution. (2) A gastric bal- 

 loon can be used which has been spe- 

 cially designed to cause gentle friction 

 of the gastric mucosa, thereby increas- 

 ing exfoliation. The balloon and the 

 technique for its use in obtaining gas- 

 tric specimens have been described by 

 Panico, Papanicolaou and Cooper 

 (Abrasive balloon for exfoliation of 

 gastric cancer cells, J. A. M. A., 1950, 

 143, 1308-11). Gastric specimens must 

 be fixed immediately in an equal volume 

 of 95% alcohol to prevent further de- 

 generation and digestion of cells. 



Duodenal drainage specimens are ob- 

 tained through a Rehfuss tube, allow- 

 ing the tip of it to pass from three to 

 six inches beyond the pjdorus. The 

 specimen is immediately mixed with 

 an equal volume of 95% alcohol. 



Rectal or colonic washings: In order 

 to obtain specimens which are as free 

 as possible of fecal material, the patient 

 is given a laxative at night, and a hot 

 soapsuds enema the next morning. 

 Saline washings of the rectum or colon 

 are collected two to five hours later and 

 mixed immediately with 95% alcohol. 

 A special suction apparatus for rectal 

 washings is described by Loeb and 

 Scapier (Rectal washings, technic for 

 cytologic study of rectosigmoid. Am. 

 J. Surg., 1951, 81, 298-302). Colonic 

 washings can be obrained by high 

 colonic irrigation, using normal saline. 



Preparation of smears: All specimens 

 from the gastro-intestinal tract must 

 be processed with the least possible 

 delay because of deterioration of the 

 cells. This is particularly important 

 in gastric and duodenal specimens 

 due to the action of the digestive juices. 

 The specimens are centrifuged as soon 

 as received and the supernatant fluid 

 promptly decanted. The addition of 



ten drops of Mayer's albumin per 50 

 cc. of specimen before centrifugation 

 will facilitate sedimentation. If smears 

 cannot be prepared immediately, the 

 sediment can be covered with absolute 

 alcohol and kept temporarily in the 

 refrigerator. Smears are prepared in 

 the same manner as urine smears but 

 stained by method III. 



Other Sources. 



Exudates, pleural, peritoneal, and 

 pericardial. Fifty cc. of 50% alcohol 

 is added immediately to 50 cc. of the 

 exudate and smears prepared from the 

 sediment in the same way as urine sedi- 

 ment smears, but stained by method 

 III. Fifty percent alcohol is used 

 rather than 95% so that there will be 

 less precipitation of protein material. 



Cystic and other fluids or washings. 

 An equal volume of 95% alcohol is 

 added immediately to the specimen and 

 smears prepared from sediment and 

 stained by method III. 



Nipple secretion smears. If secre- 

 tion can be obtained easily from the 

 nipple by very gentle pressure, a clean 

 slide is brought in contact with the 

 secretion expressed. Care should be 

 taken not to bring the slide in contact 

 with the nipple itself in order to prevent 

 the collection of an excess of epidermal 

 cells. The material is spread uniformly 

 with another slide, the smear is fixed 

 in alcohol-ether and stained by method 

 III. 



Method I. (Papanicolaou, G. N., 

 Am. J. Anat., 1933, 52: No. 13, supple- 

 ment). This is the original Papan- 

 icolaou technique for staining vaginal 

 smears in rodents and the human; it 

 can be used to advantage in endocrine 

 work and in cytologic studies of lower 

 mammals. 



1. Transfer slides from alcohol -ether 

 directly to 80% alcohol and run down 

 through 70% and 50% alcohols to 

 aq. dest. 



2. Stain in Ehrlich's hematoxylin 5 min. 



3. Rinse in aq. dest. and place in run- 

 ning tap water 15 min. 



4. Stain in 0.5% aq. eosin 3-4 min. 



5. Rinse well in tap water. 



6. Stain in 0.5% aq. waterblue 1 min. 



7. Rinse in tap water and run up 

 through 50%, 70%, 80%, 95% and 

 absolute alcohols, absolute alcohol 

 and xylol (equal parts), to xylol, 

 leaving in each solution long enough 

 to clear. 



8. Mount, without drying, with a cover 

 slip. 



Nuclei — dark purple; cytoplasm of 

 non-cornified cells — pale blue; cyto- 

 plasm of cornified cells — pink. 



