Live Cell Imaging Using ATTO-Conjugated Primary Antibodies

Cell preparation:

1. Plate cells in chosen chamber slides and grow 1-2 days in appropriate medium. Cells need to attach strongly to plate.
IMPORTANT! Some cell lines will need special coating i.e. poly-lysine of the chamber slides to aid in cell attachment. The specific type of coating needs to be determined empirically as it varies between chamber types and/or cell lines.

2. Wash cells 2-3 times with ice-cold assay buffer (PBS + 2% BSA + 0.05% NaN3).

Labeling:

3. Add ATTO-Conjugated primary antibody at the appropriate dilution in ice-cold assay buffer. Incubate 1 hr at 4°C.
4. Wash 3-5 times with assay buffer and drain well. Add buffer to cover cells and detect using a microscope.