Fig. 5.
Two color SMLM reveals co-localization of fatty acid analog puncta with the plasma membrane in fasting cells. a Sequential SMLM imaging of DII-BODIPY-C12 red puncta and a double PH domain of PLCdelta(1) fused to 3x mEos2 shows co-localization of fatty acid puncta with the plasma membrane in fasting cells. b The radial distance histogram of BODIPY-C12 red and PH2-3xmEos2 localizations from five cells (scaled to the same radius) confirms the co-localization with no detectable separation outside the localization error. The insets show magnifications of rendered SMLM images and overlap of BODIPY-C12 red puncta with the plasma membrane. c Sequential SMLM imaging of DII-BODIPY-C12 red puncta and Sec63-mEos2 shows no co-localization of fatty acid puncta with the cortical ER close to the plasma membrane. d The radial distance histogram of BODIPY-C12 red and Sec63-mEos2 localizations from five cells (scaled to the same radius) reveals a mean separation of fatty acid puncta from the ER of ~100 nm. The insets show magnifications of rendered SMLM images and the separation of BODIPY-C12 red puncta from the ER. Scalebar: 1 µm; insets 100 nm