Staining protocol for unicellular protists : Lysotracker 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 1XPBS ( 1 / 2 ) .
Resuspend cells with 1mL 1xPBS .
Add 1 / 1000 lysotracker dye : 1 uL of lysotracker for a 1mL final Volume of cells .
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 1XPBS ( 2 / 2 ) .
Wash with 1xPBS ( 1 / 2 ) .
Wash with 1xPBS ( 2 / 2 ) .
Incubate for 20 min .
Room temperature , protect from light .
Incubate for 5 minutes .
Room temperature , protect from light .
