Single-Molecule Antibody Slides For Fluorescence Microscopy

Coverslips (24 x 50 mm, #1, VWR,Catalogue Number 48404-453) were argon plasma cleaned (Ar plasma cleaner, PDC-002, Harrick Plasma) for 0h 30m 0s .

A trimmed gasket was placed on top of the slide (CultureWell^TM Reusable Gasket, 6mm diameter, Grace Bio-Labs, SKU: 103280).

Poly-L-Lysine 30µL (0.01 % w/v PLL, P4707, Sigma-Aldrich) was placed in the wells for 0h 30m 0s.

The PLL was removed, the wells washed three times with PBS (pH 7.4, 1x Gibco, Thermo Fisher Scientific, Catalogue number 10010023)

A secondary antibody of choice was added at a final concentration of 0.0002 mg/ml in PBS.

The antibodies were left in the wells for 5-10 seconds for sufficient surface density before the wells were washed three times with PBS.

PBS 30µL was left in the wells for imaging