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Research Article
Distribution of PEG-coated hollow polyelectrolyte microcapsules after introduction into the circulatory system and muscles of zebrafish
Ekaterina Borvinskaya, Anton Gurkov, Ekaterina Shchapova, Boris Baduev, Igor Meglinski, Maxim Timofeyev
Biology Open 2018 7: bio030015 doi: 10.1242/bio.030015 Published 5 January 2018

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Figures

  • Fig. 1.
    Fig. 1.

    Images and size profiles of prepared PMs-PEG containing FITC-BSA (used for investigation of distribution and stability). Median diameters are approximately 4.0 µm (A) and 2.7 µm (B). Fluorescent images of PMs-PEG are obtained in green channel.

  • Fig. 2.
    Fig. 2.

    Representative images of PMs-PEG in capillaries of zebrafish gills. (A-E) 7 days following injection of PMs-PEG into the fish kidney. (F) 1 day following retro-orbital injection. A shows side view; B shows bottom view; C-F show isolated gill arches. Fluorescent images of PMs-PEG are obtained in green channel.

  • Fig. 3.
    Fig. 3.

    Selected images of aggregates of PMs-PEG in zebrafish immediately after injection. (A-C) Gill filaments of D. rerio following retro-orbital (A,C) or intrarenal (B) injection. (D,E) Aggregates of PMs-PEG in a hepatic vessel of some individuals of D. rerio following retro-orbital injection. Fluorescent images of PMs-PEG are obtained in green channel.

  • Fig. 4.
    Fig. 4.

    Migration of PMs-PEG from the injection tract following intramuscular injection. (A) Representative image of D. rerio 14 days following the intramuscular injection of PMs-PEG (dotted line corresponds to the injection tract). The arrows indicate fluorescence at the base of the anal fin rays. (B) Changes in the shape and area of FITC fluorescence in fish muscles. The frame indicates the analyzed region (see Fig. 5); the arrow marks the needle puncture inlet. Fluorescent images of PMs-PEG are obtained in green channel.

  • Fig. 5.
    Fig. 5.

    Decrease of FITC fluorescence in fish muscles following injection of PMs-PEG with average diameters of 2.7 (n=5) and 4.0 µm (n=4). (A) Total fluorescence. (B) Fluorescence in area of injection tract, which is designated by the purple selection on the right.

  • Fig. 6.
    Fig. 6.

    Sagittal section of fish muscles of D. rerio 22 days following the intramuscular injection of PMs-PEG. (A) Transmission image, H&E stain. (B) Fluorescence image. The arrows indicate clusters of PMs-PEG surrounded by macrophages (granulomas); arrowheads indicate normal muscle tissue composed of regular muscle fibers.

  • Fig. 7.
    Fig. 7.

    Phagocytosis of PMs-PEG in fish muscles 22 days following the intramuscular injection of PMs-PEG. (A) Microsection of D. rerio muscle, H&E stain. The arrows indicate a capillary filled with erythrocytes. The arrowheads indicate macrophages loaded with PMs-PEG (cells with granulated cytoplasm) migrating along the outer walls of blood capillaries in the lymphatic ducts. (B,C,D) Macrophages engulfing PMs-PEG. Note intact microcapsules (arrows) in the cytoplasm of phagocytes. The nuclei of the cells are marked with arrowheads. Fluorescence image (D) shows dye localization inside of intact microcapsules in the macrophage cytoplasm.

  • Fig. 8.
    Fig. 8.

    Sagittal section of D. rerio 22 days after the intramuscular injection of PMs-PEG. (A) Fish tail with dorsal (upper arrow) and anal fins (lower arrow), H&E stain. The asterisks indicate the scale pockets; the arrowhead indicates the backbone. (B) Fluorescence image scaled from A shows accumulation of macrophages loaded with PMs-PEG at the base of the dorsal fin. The arrow indicates the epidermis that covers the scales. Fluorescent images of PMs-PEG are obtained in green channel. (C) Fluorescence image of clusters of phagocytes loaded with PMs-PEG migrating from the muscle tissue toward the fish skin. The asterisks indicate the scale pockets; arrowheads indicate the epidermis; red arrows indicate the scale plates. (D) Figure scaled from C showing basophilic coloration around bone (arrowhead) indicating leucocyte infiltration and inflammation process, H&E stain. (E) Figure scaled from C shows loaded phagocyte (arrow) in the epidermis of scale, recognized by the granulated cytoplasm, H&E stain.

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Research Article
Distribution of PEG-coated hollow polyelectrolyte microcapsules after introduction into the circulatory system and muscles of zebrafish
Ekaterina Borvinskaya, Anton Gurkov, Ekaterina Shchapova, Boris Baduev, Igor Meglinski, Maxim Timofeyev
Biology Open 2018 7: bio030015 doi: 10.1242/bio.030015 Published 5 January 2018
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