Staining protocol for unicellular protists : Mitotracker and Dapi
2 % PFA Fixation : Add PFA drop by drop and mixing carefully into 1mL cells for a final concentration of 2 % PFA .
Incubate cells for 4 min at room temperature .
Centrifuge at 1000g for 5 minutes .
Wash cells with 1X PBS ( 1 / 2 ) .
Resuspend cells with 1mL 1x PBS .
Add 1mM mitotracker dye .
Room temperature , protect from light .
Add 1 / 1000 DAPI dye .
Room temperature , protect from light .
Centrifuge at 1000g for 5 minutes and resuspend with 1xPBS .
Mount your slide and ready for observation !
Wash cells with 1X PBS ( 2 / 2 ) .
Incubate for 10 minutes .
Room temperature , protect from light .
Incubate for 5 minutes .
Room temperature , protect from light .
Wash with 1x PBS ( 1 / 2 ) .
Wash with 1x PBS ( 2 / 2 )
