Small human sperm vacuoles observed under high magnification are pocket-like nuclear concavities linked to chromatin condensation failure  F. Boitrelle, M. Albert, J.-M. Petit, F. Ferfouri, R. Wainer, M. Bergere, M. Bailly, F. Vialard, J. Selva  Reproductive BioMedicine Online  Volume 27, Issue 2, Pages 201-211 (August 2013) DOI: 10.1016/j.rbmo.2013.05.004 Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 1 Spermatozoa with well-condensed chromatin (A), partially noncondensed chromatin (B) and totally noncondensed chromatin (C). Aniline blue staining and ×100 objective. Bar=5μm. Reproductive BioMedicine Online 2013 27, 201-211DOI: (10.1016/j.rbmo.2013.05.004) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 2 Mean proportions of spermatozoa with acrosome reactions (green) and noncondensed chromatin (blue) for 450 vacuole-free spermatozoa (grade I) and 450 spermatozoa with multiple, small vacuoles (grade IIIa). P<0.0001, Wilcoxon–Mann–Whitney test. Error bars indicate standard errors of the mean. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Reproductive BioMedicine Online 2013 27, 201-211DOI: (10.1016/j.rbmo.2013.05.004) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 3 Three-dimensional deconvolution microscopy images. (A–D and E–H) Two morphometrically normal spermatozoa with a vacuole-free head, without (A–D) or with (E–H) acrosome reaction, observed at high magnification with differential interference contrast (A and E) and 3D-reconstructed images (B–D and F–H). (I–L and M–P) Two morphometrically normal spermatozoa with multiple, small vacuoles, without (I–L) or with (M–P) acrosome reaction, observed at high magnification with differential interference contrast and 3D-reconstructed images (J–L and N–P). (B, F, J, N) DAPI fluorescence; (C, G, K, O) Pisum sativum agglutinin fluorescence; (D, H, L, P) colocalization of fluorescent probes. Bars=1μm. Reproductive BioMedicine Online 2013 27, 201-211DOI: (10.1016/j.rbmo.2013.05.004) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 4 3D microscopy images of the nucleus of the spermatozoon shown in Figure 3M. (A) The nucleus surface (face a of the schematic cube); the white arrow shows a small vacuole. (B) Posterior view (face b) of the nuclear content; the white arrow shows a nuclear pocket (concavity) linked to a surface vacuole. (C) Right-side view (face c) of the nuclear content. (D) Sagittal section of the nucleus (face d), showing internal features; the white arrow shows a nuclear pocket linked to a surface vacuole. Bar=1μm. Reproductive BioMedicine Online 2013 27, 201-211DOI: (10.1016/j.rbmo.2013.05.004) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions