presently we have trypan blue but i think its works mainly for animal cells as we did not find any color differentiation between dead and alive algae. we are yet to try Neutral red, i would appreciate if someone can help with a protocol that could be efficient. thanks
Hey Esther
Kindly refer to this thread.
https://www.researchgate.net/post/Dyes_to_quantify_marine_microalgae
I hope it helps.
Trypan Blue is a big molecule that cannot cross cell wall, even when cells are dead.
Neutral Red is a good option as it accumulates in acid vacuoles. Accumulation supposes then that acidity is maintained in vacuoles by ATPases, thus algae are alive.
If you have fluorescence microscope you can try Propidium Iodide that stains dead cells. It is a tri-cation that cannot cross the plasma membrane of living cells due to the membrane potential and the accumulation of positive charges on the outer layer.
FDA is a hydrophobic conjugate of fluorescein that accumulates in the cytosol of cells with active esterases. Esterases cut acetate residues and release fluorescein in the cytosol. As it is a charged molecule it connat cross back plasma membrane. You can also use a the reduced form H2-FDA which is non-fluorescent and becomes fluorescent in cells producing ROS => respiring.
You can also use DiHydroRhodamine123 which is non-fluorescent, accumulates in mitochondria according to membrane potential and become fluorescent when oxidized by ROS.
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology