Multi‐colour direct STORM with red emitting carbocyanines

Single molecule‐based super‐resolution methods have become important tools to study nanoscale structures in cell biology. However, the complexity of multi‐colour applications has prevented them from being widely used amongst biologists. Direct stochastic optical reconstruction microscopy (dSTORM) offers a simple way to perform single molecule super‐resolution imaging without the need for an activator fluorophore and compatible with many conventionally used fluorophores. The search for the ideal dye pairs suitable for dual‐colour dSTORM has been compromised by the fact that fluorophores spectrally apt for dual‐colour imaging differ with respect to the optimal buffer conditions required for photoswitching and the generation of prolonged non‐fluorescent (OFF) states.