The best stain for morphological study of human seminal fluid's smears
الملخص
Objectives:There is a high need for proper evaluation of the morphological features of human sperms. The importance of this lies in the field of andrology, male fertility and in vitro fertilization. The wet smears can give rough clue about the shape of the sperms, but it is neither accurate nor reproducible. This study aimed to determine the best stain which can be used for seminal fluid cytology.
Methods: This study was conducted in Port Sudan, Red Sea State, Sudan in the period from October 2006 to September 2007. The total number of patients was 50. Samples which were collected from normospermic patients (NSP) were prepared by direct smear technique. Samples which were collected from oligospermic patients (OSP) and azoospermic patients (ASP) were prepared by direct smear technique and also by indirect smear techniques (concentration method). Smear samples were stained by freshly prepared Harris's Haematoxylin, Papanicolaou stain, May-
Grunwald Giemsa stains (MGG), supra vital stain, Giemsa stain and leishman's stain.
Results: In this study, the best stain was Harris's Haematoxylin (80% excellent for the head of sperm, 70% good for the neck, 59% excellent for the tail, 42% very good for cells in background). Harris's stain
was followed by papanicolaou stain and the third best stain was supra vital stain. MGG was better than Giemsa in staining of semen smears (75% good versus 25% good) in overall performance. The worst stain was Leishman's stain.
Conclusion: Stained smears must be used for the morphological study of semen samples. Harris's Haematoxylin is the best stain for semen cytological features. Stains which used for the semen samples should be
freshly prepared.
المراجع
2-Eliasson R. Supravital Staining of human spermatozoa. Fertil Steril 1977; 28:1257.
3. WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction: Cambridge, England: Cambridge University Press; 1992.
4. Steven A, Wilson I. The Haematoxylin and Eosin. Theory and practice of histopathological techniques. 4th edn. Bancroft JD, Steven A. New York: Churchill Livingstone, 1996; 6: 99-104.
5. Beltrami C A, fabris G, Marzola A et al. Staining of gastrin cells with Lead Haematoxylin. Histochemical Journal 1975; 7(1): 95-9.
6. Steven A, Wilson I G. The haematoxylin and eosin. Theory and practice of histological techniques. Edn.Bancroft JD, Steven A. New York: Churchill Livigstone, 1996: 99-102.
7. Cole E C. Studies in haematoxylin stain. Stain Technology 1943; 18:125-42.
8. Bales C E, Durfee C R. Cytologic technique In: Diagnostic cytology and its Histological Bases.4thend. Koss LG. Philadelphia: Lippincott 1992; 1452-1514.
9. Dinges H P, Wirnsberger G, Hofler H. Immunocyto- Chemistry in cytology comparative evaluation of different techniques: Analytical and Quantitative cytology and histology 1989; 11: 22-32.
10. Wittekind D. On the nature of Romanowsky Dye and the Romanowsky Giemsa effect. Clin Lab Haemat 1979;1:247-262.
11. Campana A, deagostini A, Bischof P. Evaluation of infertility. Hum Reprod 1995; 6: 586-606.
12. Seibel M M, Zilberstain M. The diagnosis of male infertility by semen quality the shape of sperm morphology. Hum Reprod 1995; 10: 2-247-252.
13.Ombelet W , Bosmans E, Janssen M. Semen parameters in a fertile versus subfertile population a need for change in the interpretation of semen testing. Hum Reprod 1997; 12: 987-993.
14. World Health Organization. Laboratory Manual Examination of Human Semen and Semen-Cervical Mucous interaction: 3rd ed. Cambridge University press, 1992.
15. Rajasingham S S. simplification of staining procedures in: protocols for semen analysis in clinical diagnosis. New York; the Parthenon publishing group, 2002; 3: 36.
16. Menkveld R, Stander F S, Kotze T J et al. The evaluation of morphological characteristics of human spermatozoa according to stricter criteria. Hum Reproduction 1990; 5: 586-592
