Noninvasive imaging of spindle dynamics during mammalian oocyte activation  Paula A.A.S. Navarro, M.D., Ph.D., Lin Liu, Ph.D., James R. Trimarchi, Ph.D., Rui A. Ferriani, M.D., Ph.D., David L. Keefe, M.D.  Fertility and Sterility  Volume 83, Issue 4, Pages 1197-1205 (April 2005) DOI: 10.1016/j.fertnstert.2004.07.983 Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Spindle imaging of CD-1 mouse metaphase II oocytes using the Pol-Scope (A) and fluorescence microscopy (B). Normal barrel-shaped spindles (A and B) with chromosomes aligned across the mid-region over the metaphase II plate (B) of a normal spindle. Green, meiotic II spindles stained with anti–β-tubulin and FITC-conjugated 2nd antibody; blue, chromosomes stained with Hoechst 33342. Scale bar = 10 μm. Fertility and Sterility 2005 83, 1197-1205DOI: (10.1016/j.fertnstert.2004.07.983) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Representative birefringence images of the same living CD-1 mouse oocyte, taken sequentially using the Pol-Scope, showing spindle dynamics during activation with strontium. Fertility and Sterility 2005 83, 1197-1205DOI: (10.1016/j.fertnstert.2004.07.983) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Representative birefringence images of the same living CD-1 mouse karyoplast, separated from an intact oocyte by centrifugation, taken sequentially using the Pol-scope, showing spindle dynamics during activation by strontium. Fertility and Sterility 2005 83, 1197-1205DOI: (10.1016/j.fertnstert.2004.07.983) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Representative birefringence images of the same living CD-1 mouse karyoplast, separated from an intact oocyte by centrifugation, taken sequentially using the Pol-Scope, showing spindle dynamics during activation by the protein kinase C agonist phorbol ester. Fertility and Sterility 2005 83, 1197-1205DOI: (10.1016/j.fertnstert.2004.07.983) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 5 (A) Representative birefringence images of the same living CD-1 mouse oocyte taken sequentially using the Pol-Scope, showing spindle dynamics during activation with strontium followed by administration of cytochalasin D. Cytochalasin D, an inhibitor of actin polymerization, did not disrupt spindles, prevent spindle elongation, or increase spindle birefringence during the oocyte activation process, but did inhibit spindle rotation and separation. (B) Representative birefringence images of the same living CD-1 mouse oocyte taken sequentially using the Pol-Scope, showing spindle dynamics during activation with strontium followed by administration of nocodazole (10 minutes). Nocodazole, an inhibitor of tubulin polymerization, disrupted spindles and inhibited formation of the second polar body less than 5 minutes after its administration. Fertility and Sterility 2005 83, 1197-1205DOI: (10.1016/j.fertnstert.2004.07.983) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions