Clin Exp Reprod Med.  2019 Jun;46(2):60-66. 10.5653/cerm.2019.46.2.60.

Evaluation of black glutinous rice (Oryza sativa L) extract as a novel nuclear stain for human sperm head assessment by microscopic examination

• Chomean S
• Sukanto T
• Piemsup A
• Chaiya J
• Saenguthai K
• Kaset C

• KMID: 2452644
• DOI: http://doi.org/10.5653/cerm.2019.46.2.60

Abstract

OBJECTIVE
To compare black rice (Oryza sativa L) extract with three different staining methods for human sperm head assessment.
METHODS
Semen samples were collected from 34 volunteers. Four smears of each ejaculate were prepared for staining using the rapid Papanicolaou (PAP) stain, SpermBlue, DipQuick, and black rice extract. The percentage of defective sperm heads (mean±standard deviation) was compared.
RESULTS
Black glutinous rice extract, a natural dye, was used instead of hematoxylin to stain the nuclei of the sperm heads. The percentage of defective sperm heads showed a significant difference between black rice extract and DipQuick (p=0.000). In contrast, black rice extract and rapid PAP showed no statistically significant difference (p=0.974). A strong correlation (r =0.761) was found between the findings obtained using rapid PAP and black rice extract. In contrast, a weak correlation (r =0.248) was obtained between DipQuick and black rice extract for the percentage of defective sperm heads.
CONCLUSION
The results showed good agreement and a strong correlation between the rapid PAP and black rice extract stains. The advantages of black rice extract as a novel substitute for hematoxylin for nuclear staining include ease of preparation, local availability, and favorable nuclear staining properties. Further studies could also focus on comparing staining techniques in clinical samples.

Keyword

Black glutinous rice; Microscopic examination; Papanicolaou; Rapid staining; Sperm head

MeSH Terms

Coloring Agents
Hematoxylin
Humans*
Semen
Sperm Head*
Spermatozoa*
Volunteers
Coloring Agents
Hematoxylin