Figure 5.

Development of human lacrimal glands

All panels are LSFM images of solvent-cleared eyes and heads immunostained with anti-Sox9 (A–H) and combined with MHC (A) and synaptophysin (D).

(A) Frontal view of the left eye of a PCW11.3 fetus. The orbiculari oculari muscle is seen as well as the upper (uel) and lower (lel) eye lids. The eye is pseudocolored in blue. The lacrimal gland (arrowheads) and its individual ducts (colored) are seen lying on the superolateral side of the eyeball.

(B–D) Organization the lacrimal gland in a PCW10.1 fetus. (B) shows the raw image obtained with Sox9 staining. In (C), all individual subglands are individually segmented and pseudocolored, the longest one in red. (D) shows the time course of lacrimal gland development, in situ on the eyeballs, between PCW7.5, when only a few buds are observed, and PCW11.3.

(E) All images are at the same scale to illustrate the growth of the lacrimal glands. All subglands have been individually segmented and colored. The color code is conserved, with the longest one in red as a reference (see also Figure S6).

(F) Frontal view of the face and nose/mouth of a PCW7.5 embryo stained with anti-Sox9. Overlays are 3D rendering images of the face (in gray). The superior (slc) and inferior (ilc) lacrimal canaliculi and nasolacrimal duct (ld) have been segmented and pseudocolored in orange.

(G–H) The developing eye lashes are labeled with Sox9 and form 2 arrays lining the edges of the upper (uel) and lower (lel) eyelids at PCW11.3 (arrowheads in G and H). At PCW13 (H), the number of Sox9+ buds has increased and might also include the developing meibomian glands.

Related to Figure S6.

Scale bars: all panels are counted from left to right for each row; 150 μm in (A); 500 μm in (B, left panel), (D), and (E, fourth panel); 400 μm in (B, right panel), (C), and (E, all panels except for the first and fourth); 200 μm in (E, first panel); and 1 mm in (F)–(H).