Efficacy of the Aniline Blue-eosin Staining Method for Testicular Sperm Chromatin Condensation Assay in Azoospermia.
- Author:
Yong Seog PARK
1
;
Jee Heun SHON
;
Joong Shik LEE
;
Ju Tae SEO
Author Information
1. Laboratory of Reproductive Biology & Infertility, Cheil General Hospital & Women's Healthcare Center, Kwandong University College of Medicine, Gangneung, Korea.
- Publication Type:Original Article
- Keywords:
Aniline blue-eosin (AB-E) staining;
Azoospermia;
Chromatin condensation;
Testicular sperm
- MeSH:
Aniline Compounds;
Azoospermia;
Biopsy;
Cell Count;
Chromatin;
Eosine Yellowish-(YS);
Freezing;
Histones;
Humans;
Sperm Head;
Spermatozoa
- From:Korean Journal of Andrology
2010;28(2):107-111
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: This study was performed to evaluate chromatin condensation of morphologically mature sperm using a modified aniline blue-eosin (AB-E) staining method in azoospermia. MATERIALS AND METHODS: Chromatin condensation was analyzed using an AB-E staining method in 61 cases (50 patients) of TESE or testicular biopsy with the patient's own sperm. Obstructive azoospermia (OA) was present in 48 cases in 39 patients and non-obstructive azoospermia (NOA) was present in 13 cases in 11 patients, respectively. Immature sperm heads were stained dark blue, whereas mature sperm were stained red-pink by the eosin. The percentage of sperm chromatin condensation was calculated from the ratio of the number of red-pink sperm to the total number of sperm analyzed. RESULTS: The percentage of chromatin maturity was 37.7% vs. 30.3% in OA and NOA, respectively, of the total sperm cell count. The maturity of fresh testicular sperm was 38.3% and 36.3% in OA and NOA, respectively. Also, the maturity of thawed testicular sperm was 34.5% and 10.3% (p<0.05) in OA and NOA, respectively. The maturity of fresh and thawed testicular sperm was 36.3% and 10.3% (p<0.05), respectively, in NOA. These results suggest that chromatin condensation is less stable in sperm of NOA and freezing and thawing procedures may impair sperm chromatin condensation. CONCLUSIONS: In our results, the aniline blue-eosin staining method improved the visualization of excessive histones in sperm and the diagnosis of sperm immaturity in morphologically normal testicular sperm. We found that AB-E staining method can be an effective method for analyzing testicular sperm chromatin condensation in azoospermia.
