31
Cytopreparatory Techniques
Table 31.7
Nongynecologic Staining Procedures
University of Rochester cytopathology laboratory
Memorial Sloan-Kettering Cancer Center
Fixation
95% ethanol
15 min
95% ethanol
20 min
Hydration
50% ethanol
10 dips
50% ethanol
30 s
Running water bath
30 s to 1
min
Tap water
30 s
Nuclear stain
Hematoxylin #2
(Richard Allan)
50 s
Gill I hematoxylin
(progressive)
2% min
(may vary with stain)
Blueing
Running water bath
30 s to 1
min
Running tap water
1
min
Clarifier #2 (Richard Allan)
2 quick dips—30 s
Scott's tap water
1
min
Running water bath
30 s to 1
min
Tap water
20 s (20 dips)
Blueing reagent (Richard Allan)
30 s to 1
min
Dehydration
50% ethanol
10 dips
50% ethanol
30 s (30 dips)
95% ethanol
10 dips
95% ethanol
30 s (30 dips)
Cytoplasmic stain
Orange G (Harleco)
1
min 15 s
OG-6
2 min
Rinse
95% ethanol
10 dips
95% ethanol
30 s (30 dips)
95% ethanol
10 dips
95% ethanol
30 s (30 dips)
95% ethanol
30 s (30 dips)
Cytoplasmic and nucleolar
stain (RNA related)
Eosin (EA) 65 (Harleco)
3 min
EA modified
9 min
Rinse
95% ethanol
10 dips
95% ethanol
30 s (30 dips)
95% ethanol
10 dips
95% ethanol
30 s (30 dips)
95% ethanol
30 s (30 dips)
Dehydration
99% ethanol
10 dips
100% ethanol
30 s (30 dips)
99% ethanol
10 dips
100% ethanol
30 s (30 dips)
99% ethanol
10dips
100% ethanol
30 s (30 dips)
Clearing
Xylene
10 dips
Xylene
5 dips
Xylene
10 dips
Xylene
5 dips
Xylene
10 dips
Xylene
5 dips
Xylene
15 min
Less tim e is required to prepare sta in in g s o lu tio n , since
ready-m ade products th a t produce reliab le and consistent
staining are available fo r purchase.
M an ua l staining procedures are s till p erform ed in m any
laboratories, especially w h en dealing w ith p o te n tia lly infectious
cell samples and w ith cell samples th a t m ay contam inate stain-
ing solutions. So w h ile new m ethod ologies m ay have replaced
previous preparatory m ethods, it is s till necessary to learn the
basic principles o f cell fix a tio n , staining, cover slipping, and cell
preparation.
References
1.
Allen K.
A Guide to Cytopreparation.
Dur-
ham: ASCT, 1998.
2.
Bales CE, Durfee GR. Cytologic techniques.
In: Koss LG (ed.)
Diagnostic Cytopathology
and Its Histologic Bases
.
3rd edn. Philadel-
phia: Lippincott, 1979: 1187-1266.
3.
Gatscha RM. Current concepts in cyto-
preparation and laboratory safety. In:
Schmidt WA, M iller TR, Katz RL (eds)
Cytopathology Annual
.
Chicago: ASCP
Press, 1994: 29-30.
4.
Holmquist MD, Keebler CM. Cytoprepara-
tory techniques. In: Keebler CM, Reagan JW
(eds)
The Manual of Cytotechnology
. 6th edn.
Chicago: ASCP Press, 1983: 363, 411-448.
5.
Keebler CM, Reagan JW, Wied GL.
Compendium on Cytopreparatory Techniques
.
4th edn. Chicago: Tutorials o f Cytology,
1976.
6.
Smith MJ, Naylor B.
Techniques of Cytopa-
thology: Laboratory Manual
.
Ann Arbor:
University o f Michigan,
1970: 8-10.
7.
Bibbo M, Bartels PH, Bahr GF, et al.
Computer recognition o f cell nuclei
from the uterine cervix.
Acta Cytol
1973;17:340-350.
8.
Bishop JW, Marshall CJ, Bentz JS.
New technologies in gynecologic cytol-
ogy.
J Reprod Med
2000;45:701-719.
9.
Hutchinson ML, Isenstein LM, Gold-
man A, et al. Homogeneous sampling
accounts for the increased diagnostic
accuracy using ThinPrep processor.
Am J
Clin Pathol
1994;101:215-219.
10. Hutchinson ML, Zahniser DJ, Sherman
ME, et al. U tility o f liquid-based cytology
for cervical carcinoma screening: results
o f a population-based study conducted
in a region o f Costa Rica w ith a high
incidence o f cervical carcinoma.
Cancer
(Cancer Cytopathol)
1999;87:48-55.
11. Kurtycz DF, Hoerl HD. Thin-layer tech-
nology: tempered enthusiasm.
Diagn
Cytopath
2000;23:1-5.
1001
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