- Title
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Beyond the whole-mount phenotype: high-resolution imaging in fluorescence-based applications on zebrafish
- Authors
- Oralová, V., Rosa, J.T., Soenens, M., Bek, J.W., Willaert, A., Witten, P.E., Huysseune, A.
- Source
- Full text @ Biol. Open
Imaging of transgenic zebrafish lines. (A,D,G) Whole-mount, (B,E,H) low- and (C,F,I) high-magnification (B,E,F,H,I) cross sections and (C) sagittal section. Lines in A,D,G indicate approximate level of sectioning in B,E,H and C,F,I. (A–C) Tg(sox17:egfp) zebrafish with GFP-positive endodermal cells along the midline, extending into the pouches (arrows). Blood vessels also show a fluorescent signal. (D–F) Tg(krt4:gfp) zebrafish with GFP-positive periderm (arrows). Labeled cells also cover the oropharyngeal lining (arrowheads). (G–I) Et(Gal4-VP16)zc1044A;Tg(UAS-E1b:nsfB-mCherry)c264, abbreviated as GET-periderm line (GET, Gal4 enhancer trap) (Eisenhoffer et al., 2017). The periderm is labeled red (arrows). b, brain; e, eye; nt, notochord; ov, otic vesicle; y, yolk. Scale bars: (A,D) 250 µm; (E,G,H) 100 µm; (B,C,F,I) 50 µm. |
Imaging of photoactivatable kaede. Vertebral end plates of an adult Tg(osx:kaede) zebrafish shown using a GFP (A–C) and rhodamine (A′–C′) filter, before (A,A′), after 5 s (B,B′) and 30 s (C,C′) of exposure to a wavelength of 365 nm. Note the fading of the green signal from A to C and strengthening of the red signal from A′ to C′ in the osteoblasts lining the vertebral end plates (arrowheads). All pictures were taken using the same exposure time (1.6 s). Scale bar: 25 µm. |
Whole-mount immunostaining. (A,D,G) Whole-mount, (B,E,H) low- and (C,F,I) high-magnification cross sections. Lines in A,D,G indicate approximate level of sectioning in B,E,H and C,F,I. (A–C) Wild-type (WT) zebrafish immunostained for laminin. Note the basal lamina delimiting optic cup, brain, epidermis and endodermal pouches. (D–F) Tg(sox17:egfp) zebrafish pulse-labeled with BrdU and whole-mount stained with an anti-BrdU antibody. (G-I) Tg(sox17:egfp) zebrafish immunostained with a pan-cytokeratin antibody. The whole-mount images in D,G show the red channel only; the GFP fluorescence is likewise visible prior to dehydration and embedding. b, brain; e, eye; ep, epidermis; oc, optic cup; p, endodermal pouch. Scale bars: (A,D,G) 250 µm; (E,H) 100 µm; (B,F,I) 50 µm; (C) 25 µm. |
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