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. 2020 Jul 20;11:3638. doi: 10.1038/s41467-020-17360-9

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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. 7 — a, b Schematics showing the site of microinjection within a 2 dpf embryo and the evolving fluorescence of pHrodo-containing DOPC:4 → 3 liposomes within the tail region of a wildtype (AB/TL) embryonic zebrafish, over time. The fluorescence intensity of pHrodo increases >100-fold in mildly acidic environments (e.g. late endosomes/lysosomes, pH < 6) and is, therefore, particularly apparent within SECs—cells with exceptionally high lysosomal activity. c Tissue level views of evolving pHrodo-associated fluorescence over time either in the absence of UV irradiation and following in situ UV irradiation (15 min, 370 ± 7 nm, 2.4 J cm⁻²). In the absence of UV irradiation, pHrodo-associated fluorescence is observed within a small number of cells within the CHT of the embryo (white arrowheads). Liposomes contained 1 mol% DOPE-pHrodo for visualisation. Scale bar: 50 μm. d CryoTEM images of SR-B filled, DOPC:4 liposomes before and after in situ irradiation (15 min, 370 ± 7 nm, 202 mW cm⁻²). Scale bars: 100 nm. e Maximum intensity projections of two-photon z-stacks (spanning the full width of the embryo) showing SR-B filled DOPC:4 liposome distribution, before and after UV irradiation. Scale bar: 50 μm. f Time-lapse images of SR-B filled DOPC: 4 → 3 liposome distribution during UV irradiation. Scale bar: 50 μm. g Mean SR-B fluorescence intensity within ROIs (lumen of the DA, orange line; DA vessel wall, green line, shown in e), before and during UV irradiation. SR-B fluorescence intensity in circulation decreases upon UV irradiation with a concomitant increase in SR-B fluorescence intensity associated with the DA blood vessel wall. Liposomes (e–g) containing encapsulated SR-B (10 mM) and otherwise unlabelled.