Iterative fluorescence imaging

A kit of parts (set) comprising a blocking buffer comprising:

• i. a blocking compound that is capable of binding to hydrophobic binding sites non-specifically,
• particularly wherein the blocking compound is a blocking polypeptide,
• more particularly a blocking polypeptide selected from BSA, FBS, SA, normal serum, gelatin, skim milk powder or Prionex® Reagent,
• even more particularly 0.5 to 2% of a compound selected from BSA, FBS, SA, normal serum, gelatin, skim milk powder or Prionex® Reagent,
• most particularly ca. 1% BSA and
• b. an imaging buffer comprising
• i. a radical scavenger, particularly a compound that is selected from N-acetylcysteine, N-acetylcysteine amide, cysteine, L-ascorbic acid, resveratrol, β-carotene, seleno-L-methionine, chlorogenic acid and/or caffeic acid, more particularly 1 mmol/L to 1 mol/L of a compound selected from N-acetylcysteine, N-acetylcysteine amide, cysteine, L-ascorbic acid, resveratrol, β-carotene, seleno-L-methionine, chlorogenic acid or caffeic acid, most particularly 700 mM N-acetylcysteine and
• ii. a pH of 7.2-7.6, particularly a pH of 7.4.
• c. an elution buffer comprising
• i. a reducing agent, particularly a compound that is selected from TCEP, dithiothreitol, or 2-mercaptoethanol, more particularly 1 mmol/L to 1 mol/L of a compound that is selected from TCEP, dithiothreitol, or 2-mercaptoethanol, most particularly 70 mM TCEP and
• ii. at least one compound disrupting hydrogen bonds, particularly a chaotropic salt, particularly a compound that is selected from guanidinium chloride, urea, sodium dodecyl sulfate or magnesium chloride, more particularly 1 mmol/L to 10 mol/L of one or more of guanidinium chloride, urea, sodium dodecyl sulfate or magnesium chloride, most particularly 3 M guanidinium chloride and 3 M urea and
• iii. a buffering component, particularly L-glycine, more particularly 0.5 M L-glycine and
• iv. a pH lower than (<) 4, particularly pH <3, more particularly <2.7.