Fig. 2: Phasor-FSTM for two-color super-resolution imaging of a living HeLa cell. | Light: Science & Applications

Fig. 2: Phasor-FSTM for two-color super-resolution imaging of a living HeLa cell.

From: Phasor-FSTM: a new paradigm for multicolor super-resolution imaging of living cells based on fluorescence modulation and lifetime multiplexing

Fig. 2: Phasor-FSTM for two-color super-resolution imaging of a living HeLa cell.

a Intensity image of lysosomes and mitochondria composed of the photons in all time channels. b Fluorescence decay curve of the obtained FLIM image. c Gaussian and donut images obtained by temporal modulation. Inset: phasor plots and photon extraction using phasor analysis. d Intensity unmixing by phasor analysis to decompose the structures labeled with two dyes and excited with Gaussian and donut laser beams, and spatial modulation to achieve super-resolution (FSTM). e Phasor-FSTM (two-color super-resolution) image by overlapping two color-coded FSTM images in d. f Magnified views of the white squares in confocal, FSTM, and Phasor-FSTM images in c–e. g Intensity profiles along the arrows in f. h Object identification using adaptive thresholding on the images of Phasor-Confocal and Phasor-FSTM. i 2D morphological analysis of mitochondria and lysosomes in confocal and FSTM images. Data are presented as the means ± SE. j Example of common mitochondrial network. Networks are mitochondrial structures with at last a single node and three branches. Scale bars, 5 μm (a, c–e, h) and 500 nm (f, j)

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