We have all been there, after spending a day or two fixing and staining your samples, the images don't look as good as expected, the signal is not optimal, and no obvious conclusion can be drawn from the data. We have put together some tips to help you optimize your immunofluorescence staining protocol and increase the signal-to-noise ratio in your samples. If you are unsure about how immunofluorescence works, we recommend you read our techniques page first.
When preparing a sample for STORM microscopy, one of the most important factors to consider is the choice of fluorophore. The ability to accurately localize single molecules is dependent on the use of fluorescent molecules that stochastically switch between a fluorescent “on” state and a dark “off” state – a photophysical behavior that is often referred to as blinking.