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.

6 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?

    • Ido –
      I don’t really know. We’ve always just transferred to low fluorescence media immediately prior to imaging, but I don’t know if the background would be even lower if we did the transfer earlier.

  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.

    • Hi Joan – I’m an R&D scientist at Thermo Fisher Scientific and would be interested in hearing more about your issues with our FluoroBrite DMEM. Are you doing your overnight studies in a standard 5% CO2 incubator? FluoroBrite DMEM’s formula is based on the standard DMEM formulation and I wouldn’t expect the pH to be any different than what you see in your other culture medium. Can you tell me what cell type(s) and basal media you are currently using?


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