Figure | Excitation spectral microscopy integrates multi-target imaging and quantitative biosensing. (IMAGE)
Light Publishing Center, Changchun Institute of Optics, Fine Mechanics And Physics, CAS
Caption
a, Schematic of the setup. Full-frame spectral micrographs are obtained by the synchronized fast modulation of the excitation wavelength in consecutive frames. P, polarizer; L, lens; F, bandpass filter; DM, dichroic mirror. b, Unmixed images of 6 subcellular targets in a live COS-7 cell with 8 excitation wavelengths. LipidSpot 488: lipid droplets (LDs), SYBR Green: mitochondrial DNA, Mito-PhiYFP: mitochondrial matrix, WGA-CF532: cell membrane, LysoBrite Orange: lysosomes, tdTomato-ER3: ER. c, Reference excitation spectra of the 6 fluorophores, separately measured on the setup using singly labeled samples. d, Mito-pHRed absolute pH maps of the mitochondrial matrix in a live HeLa cell, before (top) and after (bottom) 120 s treatment with 20 μM CCCP. e, Color-coded FRET maps for a macromolecular crowding sensor, for two live COS-7 cells before (left), ~10 s after (center), and ~25 s after (right) 150% hypertonic treatment. f, Unmixed images of color-coded Mito-pHRed absolute pH map, mOrange2-Parkin, PhiYFP-LC3, and LAMP1-Clover for two Parkin-expressing live HeLa cells after the application of 20 μM CCCP for 4 h. g, Zoom-in of the white box in (f)
Credit
by Kun Chen, Rui Yan, Limin Xiang, and Ke Xu
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