SB1969 (wildtype)

 

1) grow log cells.

2) add 25 microliters of Mito-Green to 25 mLs of cells for one hour.

3) wash out and starve, for 2 hrs.

4) label with DAPI (1 microliter per mL) and FM4-64 (10 microliters per mL)

            five minutes (max)  {do 2 mLs at a time}

5) Wash out 3 X.  Immobilize, photograph.

 

 

Anti-cDNA12 (lumpy mutant)

1) grow log cells.

2) starve, for 2 hrs.

3) label with DAPI (1 microliter per mL) and FM4-64 (10 microliters per mL)

            five minutes (max)  {do 2 mLs at a time}

5) Wash out 3 X.  Immobilize, photograph.

 

 

Three variations: 

 

1) Take photos at 5-minute intervals (independently).

 

2) Mix cells/ label simultaneously.  Attempt to photograph two-cells at a time.   (+/-)

 

3) Follow one cell (each) with confocal over 15 minutes or so generating time course.