Fluorescent staining of lysosomes and mitochondria?

Question

Why use NaOH?

Answer 1

Do you mean NAO?
The hydrophobic fluorescence dye 10-n-nonyl-acridinium-orange-chloride, NAO, stains specifically the mitochondria of living HeLa-cells. A dye concentration of 1 X 10(-8) M is sufficient for vital staining and at 5 X 10(-7) M an incubation time less than 1 min is enough to generate the bright green fluorescence of the mitochondria. The retention of NAO by the mitochondria is longer than 7 days. The dye accumulation is not affected by the ionophores valinomycin, nigericin, gramicidin, the uncoupling agents DNP, CCCP or by ouabain. In contrast to Rh 123 the trans-membrane potential is not the driving force of the NAO accumulation. We assume that NAO is bound to the hydrophobic lipids and proteins in the mitochondrial membranes by hydrophobic interaction.
Sorry if this isn't what you meant!

Answer 2

Or did you mean MeOH. I know when you label lysosomes and use something like a mono-clonal LAMP anti body that you need to methanol fix because PFA doesn't work.

Answer 3

isnt it to make the membrane permiable? im sure thats why i used it in my practicles in the first year of uni...Antibody staining is different although you still have to make the cell membrane permiable.