PAPANICOLAOU TECHNIQUES 



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PAPANICOLAOU TECHNIQUES 



tion of more specific nature but, unless 

 prepared and fixed very quickly, swab 

 smears are apt to be drier than aspira- 

 tion smears and therefore diSicult to 

 interpret. 



Smears of the female genital tract 

 are stained by either method I or II. 



Urinary Tract. 



Collection of specimens: Urine from 

 females must be catheterized to avoid 

 contamination from vaginal secretion. 

 Ureteral specimens are advisable if 

 carcinoma of the kidney pelvis is sus- 

 pected. In cases of suspected carci- 

 noma of the prostate, separate voided 

 urine specimens are obtained before 

 and after massage. In such cases, 

 prostatic smears can also be prepared 

 by spreading prostatic secretion di- 

 rectly on slides and fixing them in alco- 

 hol-ether. 



Preparation of smears: Upon collec- 

 tion, the urine specimen (approximately 

 50 cc. of a bladder specimen; as much 

 of a ureteral specimen as can be ob- 

 tained) must be mixed immediately 

 with an equal volume of 95% alcohol 

 for fixation and preservation of the 

 cells. The mixture of urine and alcohol 

 is centrifuged in 50 cc. conical tubes at 

 medium speed for 15-30 minutes (until 

 sediment is well packed), after which 

 the supernatant liquid is decanted. 

 Clean slides, marked in advance with 

 a diamond pencil or water-proof lab- 

 oratory ink, are coated with a thin film 

 of Mayer's albumin fixative in order to 

 make the cells adhere to the slide. 

 Sediment can be removed from the 

 tube by means of a small spoon-shaped 

 instrument such as a nasal or small 

 bone curette or, if very scanty, by 

 pipetting. It is spread on the slide, 

 using a second clean slide to distribute 

 it in a uniform smear. When the smear 

 starts to dry at its periphery, it is im- 

 mersed in 95% alcohol and ether, 

 equal parts. Care must be taken, 

 however, not to allow the smear to 

 become too dry before fixation for the 

 reasons discussed previously. The 

 smears should be left in the alcohol- 

 ether fixative for at least one hour 

 before staining or they tend to wash 

 off during the staining process. All 

 instruments used in transferring sedi- 

 ment from tube to slide must be care- 

 fully cleaned to avoid contamination 

 of the next specimen. If there is suffi- 

 cient sediment, three or four smears 

 are prepared from each specimen. If 

 there is an abundance of sediment, it 

 should be stirred up in the tube in 

 order to insure representative sampling. 

 If smears cannot be prepared imme- 

 diately after centrifugation, the super- 



natant liquid is decanted, the sediment 

 covered with a few cubic centimeters 

 of absolute alcohol and placed in the 

 refrigerator. 



Graham et al. (Vincent Memorial 

 Hospital, The Cytologic Diagnosis of 

 Cancer. W. B. Saunders Co., 1950) 

 suggest mixing two or three drops of 

 albumin fixative with the sediment in 

 the centrifuge tube before making 

 smears so that the cells will stick better 

 to the slides. This has been found to 

 be very helpful in urine as well as 

 other sediments. 



Urine smears are stained by method 

 IV. 



Respiratory Tract. 



Sputum: This must be a deep cough 

 specimen as a salivary specimen is 

 obviously of no value in the diag- 

 nosis of carcinoma of the lung. It 

 is ordinarily collected directly in 70% 

 alcohol to retard the growth of bacteria 

 and fungi as well as to prevent cellular 

 degeneration. However, if smears can 

 be made within four hours after the 

 specimen is obtained, it need not be 

 fixed in alcohol. Since alcohol hardens 

 sputum, smears are more easily pre- 

 pared from a fresh, unfixed specimen. 

 When alcohol is used for fixation, 70% 

 is preferable to 95% because it causes 

 less hardening of the sputum. Before 

 preparing the smears, the specimen is 

 examined for blood-tinged portions. 

 If any are present, they are transferred 

 to slides which have been marked pre- 

 viously and coated with a thin film of 

 Mayer's albumin. If no blood tinged 

 particles are found, samples from differ- 

 ent parts of the specimen are to be 

 transferred to the slides. A fairly 

 thin uniform smear is prepared by 

 spreading the material with another 

 clean slide, gently rubbing it between 

 the two slides in order to crush any 

 hard rubbery masses and distribute 

 them evenly. The smear is fi.xed, with- 

 out drying, in alcohol-ether fixative for 

 a minimum of one hour. Three or 

 four smears should be prepared. They 

 are stained by method III. 



Bronchial aspirates or saline washings 

 of the bronchus are mi.xed immediately 

 with an equal volume of 95% alcohol. 

 Because of the scantiness of material 

 obtained by bronchial aspiration, the 

 collection tube should always be rin.sed 

 well with alcohol and the rinse solution 

 added to the specimen. Specimens are 

 centrifuged and smears prepared from 

 the sediment by the method given for 

 smears of urine sediment. They are 

 stained by method III. 



