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. 2022 Feb 26;25(4):103993. doi: 10.1016/j.isci.2022.103993

Figure 4.

Figure 4

Hyaline cells proliferate in vitro (Figures 4A–4D) and in vivo (6-h samples: Figures 4E and 4G; 7-day samples: Figures 4F and 4H)

(A) Hemolymph was withdrawn from crayfish and hemocytes were attached to lysine-coated slides. All three hemocyte types can be identified in these samples: granular (arrows); semigranular (open arrowheads); hyaline (filled arrowheads). These hemocytes were treated with BrdU for 6 h, beginning 1 h after hemolymph was taken from the crayfish, and then processed immunocytochemically. Only hyaline cells in these samples contain labeled nuclei (BrdU, red), although not all hyaline cells are labeled. Many granular cells contain a small area of cytoplasmic labeling (arrow in the middle of the frame; see also B, D).

(B) Two hyaline cells containing nuclear BrdU labeling are shown, as well as one granular cell with an area of cytoplasmic labeling (arrow). Two unlabeled hyaline cells are also shown in the bottom right of this image.

(C and D) Higher magnification images are shown with (C) two hyaline cells containing nuclear labeling and (D) two granular cells with a small area of cytoplasmic labeling.

(E) In vivo 6 h post-BrdU injection. Hemolymph was withdrawn from crayfish that had been injected with BrdU 6 h earlier. Hemocytes were attached to lysine-coated slides, fixed and processed immunocytochemically. All three hemocyte types can be identified in these samples: granular (arrows); semigranular (open arrowheads); hyaline (filled arrowheads). Only hyaline cells in these samples contain labeled nuclei (BrdU, red), although not all hyaline cells are labeled.

(F) In vivo 7 days post-BrdU injection. Hemolymph was withdrawn from crayfish that had been injected with BrdU 7 days earlier, and hemocytes were attached to lysine-coated slides. All three hemocyte types could be identified in these samples, although only hyaline cell nuclei contain BrdU labeling. Examples of hyaline cells, two of which are labeled, and semigranular cells, which are not labeled, are illustrated.

(G) In vivo 6 h post-BrdU injection. Higher magnification, showing a BrdU-labeled hyaline cell (filled arrowhead) near unlabeled hemocytes.

(H) In vivo 7 days post-BrdU injection. Higher magnification, showing BrdU-labeled hyaline cells amidst unlabeled cells. Scale bars: (A) 20 μm; (B–H) 10 μm.