Fluorobrite DMEM and sptPALM

A number of our users have been testing out the low-fluorescence DMEM from Life Technologies, Fluorobrite DMEM, and many have found that it substantially improves the signal-to-background of their experiments, particularly for imaging dim samples. Recently we tried it when doing sptPALM of PA-GFP, and found that it made a substantial difference in image quality. Below are two movies, taken under identical conditions, and scaled identically, of single PA-GFP labeled receptors diffusing in the membrane of a cell. They are illuminated with modest 488 nm excitation and weak 405 nm excitation to switch PA-GFP on. The movies were acquired by TIRF, and are shown in real time. The top movie is of┬ácells in conventional DMEM-based media; the bottom, cells in Fluorobrite DMEM. The Fluorobrite media substantially improves the image contrast; there’s also a noticeable reduction in background events outside of the cell.

If you’re doing low-signal imaging, particularly in the GFP channel, I think the Fluorobrite media is well worth trying out.

5 thoughts on “Fluorobrite DMEM and sptPALM

  1. Pingback: 3rd Cell Cyclist Newsletter | Sciency Things from the North

  2. Hello, and thank you for your insights.
    I have been wondering – is it important to have my cells in a low-fluorescent medium only during live image acquisition, or is it also advised to constitutively culture them in such medium for several passages?
    Thanks!

  3. We find that overnight to 24 hr growth in this media leaves us with a high percentage of dead cells and residual cell debris. We at first thought this was due to a contamination event but after extensive testing, we feel confident it is not..It could be pH; It is obviously hard to check pH.

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