A preliminary study of reactive oxygen species activity within motile Boer buck sperm using confocal laser scanning microscope (CLSM)

Siti Fatimah Ibrahim, I. Mohd Iswadi, Khairul Osman, M. Nurhazilah, M. S. Fadzilah, A. R Mohd Padzil, Srijit Das, O. Abas Mazni

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Objective. There is an association between reactive oxygen species (ROS) and DNA damage to sperm. Researchers believe that ROS is always present at the sperm's head. The variation of ROS concentration within the area has an impact on the integrity of the DNA. Materials and Methods. A study was conducted to confirm the location of ROS within a group of motile Boer buck sperm. A dual photon confocal laser scanning microscope (CLSM) with a 507 nm excitation and 529 nm emission was used on thawed cryopreserved sperms mixed with 10% polyvinyl pyrrolidone (PVP) and stained with 10 mmol/l dihydrorhodamine 123. ROS production was observed and captured under the CLSM at 400x magnification and a frame rate of 35 fps. All fluorescent images were then automatically overlaid with images obtained through Brightfield (BF) to the location of the sperm's membrane. A 3-D reconstruction of the sperm was also conducted to confirm the location of ROS activity. Results. ROS activity was present at the acrosome, midpiece one third of the sperm's tail and whole area of the sperm head. ROS in immotile sperms was also present at the acrosome, one third of sperm tail and the whole area of the head compartment. Relatively fluorescence intensity of ROS was found lower in motile Boer buck sperm as compared to the immotile Boer buck sperm. Conclusion. It is concluded that sperm viability largely depends on the ability of the sperm to confine ROS within the acrosome, midpiece and one third of the sperm's tail.

Original languageEnglish
Pages (from-to)125-128
Number of pages4
JournalClinica Terapeutica
Volume161
Issue number2
Publication statusPublished - Mar 2010

Fingerprint

Spermatozoa
Reactive Oxygen Species
Lasers
Sperm Tail
Acrosome
Sperm Head
Polyvinyls
Pyrrolidinones
Photons
DNA Damage
Fluorescence
Head
Research Personnel
Membranes
DNA

Keywords

  • Acrosome
  • Boer buck
  • DNA
  • Reactive oxygen species
  • Sperm
  • Viability

ASJC Scopus subject areas

  • Medicine(all)

Cite this

A preliminary study of reactive oxygen species activity within motile Boer buck sperm using confocal laser scanning microscope (CLSM). / Ibrahim, Siti Fatimah; Iswadi, I. Mohd; Osman, Khairul; Nurhazilah, M.; Fadzilah, M. S.; Padzil, A. R Mohd; Das, Srijit; Mazni, O. Abas.

In: Clinica Terapeutica, Vol. 161, No. 2, 03.2010, p. 125-128.

Research output: Contribution to journalArticle

Ibrahim, Siti Fatimah ; Iswadi, I. Mohd ; Osman, Khairul ; Nurhazilah, M. ; Fadzilah, M. S. ; Padzil, A. R Mohd ; Das, Srijit ; Mazni, O. Abas. / A preliminary study of reactive oxygen species activity within motile Boer buck sperm using confocal laser scanning microscope (CLSM). In: Clinica Terapeutica. 2010 ; Vol. 161, No. 2. pp. 125-128.
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