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. 2020 May 18;11:2460. doi: 10.1038/s41467-020-16297-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Schematic illustration of the nanogel functionalization strategy and post-polymerization modification strategy for mitochondria tracking. b Confocal images of co-localized nanogel (TPP-N6, green channel) with MitoTracker Red (Red channel) in live A549 cells. Green channel: the ring-opened form of SP (λ_ex. = 405 nm); Red channel: MitoTracker Red (λ_ex. = 630 nm). The intensity correlation plots of TPP-N6 and MitoTracker Red were processed on imageJ and Pearson correlation coefficient was analyzed using Coloc2 plugins; c Schematic illustration of the lifetime amplification strategy using the reversible method compared with the previous method which required extra-long lifetime. d FLIM images of the nanogel with mitochondria tracker (TPP-N6) in live A549 cells under the stimulation of remote UV and Vis manipulation (Irradiation time: 120 s). Similar images were obtained for more than two sets of experiments. The insert images are the lifetime histograms of the nanogel in the whole image; Scale bar: 60 μm; Color bar: Fast lifetimes (ns). e The FLIM image (top) of TPP-N6 in live A549 cells before UV irradiation was subtracted with the same range after UV irradiation calculated by ImageJ. The subtracted image (top) was compared with the confocal fluorescence (bottom) of MitoTracker Red (Red channel).