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Microscopic analysis of MTT stained boar sperm cells
1Barex Biochemical Products, Seb. Centenweg 45, 1602ML Enkhuizen, The Netherlands.
Open Veterinary Journal
|December 2, 2015
Summary
The MTT assay visualizes formazan in boar sperm, revealing its uptake by lipid droplets. This microscopic study suggests the MTT assay is qualitative and has limited practical use for sperm viability testing at artificial insemination stations.
Area of Science:
- Reproductive Biology
- Cell Biology
- Biochemistry
Background:
- The MTT assay, based on formazan production by sperm cells, has been used for mammalian sperm viability.
- Microscopic visualization of formazan localization in boar sperm was previously lacking.
Purpose of the Study:
- To microscopically visualize the precise location of formazan within boar sperm cells during the MTT assay.
- To understand the morphological events of MTT staining in boar sperm.
Main Methods:
- Boar sperm cells were incubated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide).
- Microscopic observation documented the morphological changes and formazan granule formation over time.
- Sperm cells were analyzed for formazan localization and crystal formation.
Main Results:
- MTT staining initially showed diffuse midpiece staining, progressing to intense granules.
- A distinct formazan granule appeared on the sperm head, increasing in size.
- Formazan granules disappeared, forming aggregated crystal threads in the medium, indicating uptake by lipid droplets.
Conclusions:
- Formazan is localized within lipid droplets in the cytoplasm of boar sperm.
- The MTT assay for sperm viability should be considered qualitative.
- The practical application of the MTT assay at artificial insemination stations is limited due to its qualitative nature.
