Figure 1.

Schematic of the PEGylation protocol and preparation of sample chamber for smFRET experiment

(A) Stepwise representation of surface passivation. (i) The slide with hole is properly cleaned. (ii) KOH treatment introduces the hydroxyl group (-OH) on the slide surface. (iii) The slide is ready for amino-silanization followed by PEGylation (iv).

(B) Stepwise representation of slide assembly. (i) Slice of a double sided tape applied to the PEGylated slide surface in-between each holes. (ii) Sandwich the coverslip on the taped slide where PEGylated surface facing each other. Seal the tape with pipet tip and applying quick-drying epoxy at the two edges. (iii) Sample reservoir made with a cut piece of the pipet tip (beside contain) in one hole and epoxy mold created in other hole for the syringe with tube attachment.