Lysotracker om Coprinopsis cinerea - (Jan/03/2008 )

Hello.
i am studying endocytosis in coprinopsis cinerea (a mushroom), i tried using the Lysotracker Red to label the vacuoles-like compartment in the mycelium. The observations are new and strange to me. I could see some hollow circular structure, with only the membrane stained red, but at the same time , the lumen of some of the big circular organelles are also stained completely red. I would like to ask. whether lysotrakcer Red stain only the membrane of the acidic organelle or the entire lumen, or both??

Thank you very much.

I'm no mushroom scientist (i'm doin research in the lysosome-business) but in in eucaryotes, lysostracker aggregates as a weakly basic amine in acidic comparments like the lysosome and late endosomes. The weak base is transported into those compartments and protonated in there. This protonated molecule is not able to diffuse thou membranes and is trapped.
usually the wohle lumen is stained, but you often see a stronger staining near membranes.

THank you for your suggestions and comments....

another question is does the staining ablility of the lysotracker have sth do with the age of the cells? coz i found that in younger cells, the membrane of the vacuole-like compartment are more preferentially stained wherease in older cells the more circular compartemtns get their lumen stained completely red rather than the membranes.


and, frankly i duno the pK value of it....sorry about that....and what's the importance of the pK value in this case??...


thank you very much.,!!!

could it be that the endocytotic activity and intracellulary transport differs in your cells of different ages ? or may be the pH in the vacuoles is increasing by age... ?

i think franky asked for pK value to make sure, that the vacuoles of your fungi are acidic....