Skip to main content
. 2021 Jul 26;24(8):102905. doi: 10.1016/j.isci.2021.102905

Figure 4.

Figure 4

High-throughput 3D volumetric analysis in retinoblastoma models

(A) 3D reconstruction and optical section of dispase treated, depigmented, cleared, EdU-labeled and RedDot2 nuclear-stained 1-month-old ⍺-Cre; Rbloxp/loxP; p107−/− mouse eye.

(B–D) (B and C) distribution and (D) density analysis of EdU-labeled cells in 1-month-old ⍺-Cre; Rbloxp/loxP; p107−/− mouse eyes. The EdU-labeled cells are distributed in the periphery with a higher density in the anterior-ventral-temporal quadrant of the retina. Surviving, structurally intact retina can be distinguished morphologically from structurally disrupted, ectopically dividing EdU+ cells (A inset). Solid black lines represent the outline of the retinal cup and position of the optic nerve (ON). Dashed black lines represent the outline of the iris of the eye.

(E and F) (E) 3D reconstruction and (F) optical section of an eye from a 6-month-old α-Cre; Rbloxp/loxP; p107−/−; p27+/− mouse stained with EdU. The ectopically dividing peripheral retina (dashed line) displays a higher density of EdU+ cells in the central portion of the eye with reduced density closer to the periphery.

(G) Comparison of the volume of EdU+ regions in the two retinoblastoma genotypes. Data are represented as means ± SEM, Unpaired ttest, ∗∗ denotes p < 0.01, p = 0.0032; n = 2 eyes (α-Cre; Rbloxp/loxP; p107−/−; p27+/−) and n = 3 eyes (⍺-Cre; Rbloxp/loxP; p107−/− mouse eyes). Scale bars = 500 μm. D, dorsal; V, ventral; N, nasal; T, temporal. Solid lines in the (A) 3D reconstruction and (B) distribution plots represent the plane of the image toward the reader (out of the page), while dashed lines represent the plane of the image away from the reader (into the page).

HHS Vulnerability Disclosure