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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Biophotonics. 2011 Oct 11;5(1):67–84. doi: 10.1002/jbio.201100066

Figure 7.

Figure 7

Linear unmixing was applied to flat-field corrected spectral images. (A) Total fluorescence intensity of a 10 μm-thick lung cryoslice containing perfused GFP-expressing pulmonary microvascular endothelial cells (PMVECs); (B) image of Nuclei (Hoechst 33342 staining) with 360/40 nm excitation and 450 nm emission; (C) brightfield image at 500 nm; unmixed images of autofluorescence (D), GFP (E), and background (F); the spectra used for unmixing (G) were acquired from GFP-expressing PMVECs and negative control tissue; (H) false-colored overlay of unmixed image showing autofluorescence (red), GFP (green), nuclei (blue), and background (orange). Control lung cryoslices showed minimal false-positive GFP signal with identical imaging and linear unmixing (data not shown).