Fig. 4.
Preparing the sample for imaging. (A) The slide or coverslip is functionalized with a biotin-containing moiety such as biotin-PEG or biotin-BSA. (B) An avidin family protein such as streptavidin is added to provide an anchor point for CP immobilization. (C) The CP is added and allowed to bind to streptavidin to permit capture of the target. (D) The target is added and incubated to allow capture at the surface. (E) The FP is added in an imaging solution containing an oxygen scavenger system to permit single-molecule kinetic fingerprinting through repetitive binding of the FP to the target.