Fig. 2.

FLIPPER-body labeling of nuclear localized GFP. HEK293T cells expressing H2B-GFP. a H2B-GFP was detected by indirect labeling with rabbit anti-GFP and secondary IgG conjugated to QD655. Note that fluorescence detection of QDs is limited to the cytoplasm, which may be non-specific labeling. High-resolution EM images also show only QDs in the cytoplasm. b H2B-GFP was detected by direct labeling with FLIPPER-bodies containing mCherry. The used FLIPPER-body is shown at the right. Note the specificity of the mCherry signal in only the GFP positive cells. With EM, the black DAB precipitates are seen in the nuclei. DIC differential interference contrast, GFP GFP fluorescence, QD655 QD655 fluorescence, mCherry FLIPPER-body, merged GFP and mCherry, EM ultrathin EM section. Bars LM and EM 10 µm, EM zoom in 1 µm. Unbiased large-scale high-resolution EM images are available via http://www.nanotomy.org