Ovary microscopy

Since our backbone 2 aPNA incorporates six Lys residues in its peptide sequence and is cationic at a physiological pH, we were optimistic that this aPNA would be taken up into cells without the need for any external carrier system. To answer the simple question of whether b2 aPNAs are intemahzed, a standard fluorescence microscopy experiment was performed to see if whole cells that were incubated with a fluorescent-labeled aPNA would internahze labeled material [70]. Chinese Hamster Ovary (CHO) cells in culture were incubated with BODIPY-la-beled TCCCT(b2) at 37 °C for various periods of time. Following incubation, the cells were rinsed in phosphate-buffered sahne (PBS), fixed with 4% formaldehyde at ambient temperature for 20 min, then washed with PBS and stored in a refrigerator until examined by fluorescence microscopy. 

Zucker, R.M. and Jeffay SC. Confocal Laser Scanning Microscopy of Whole Mouse Ovaries Excellent Morphology with Apoptosis Detection and Spectroscopy. Cytometry 2006 69A August 2006 930-939 (COVER). 

Fig. 7. Fine structures in the egg and around the ovary revealed by transmission electron microscopy, (a) Adjacent pronuclei in an egg filled with yolk granules. Bar = 10 pm. (b) Adjacent part of the pronuclei in (a). Bar = 5 pm. (c) The viteUine membrane (indicated by seven arrows). The envelope is still not completed, but interrupted. The vitelline membrane separates the egg with yolk granules (below) from the lyrate organ (above). Bar = 5 pm. (d) Ovary with several oocytes. Presumed sperms are visible around the ovary. Bar = 5 pm. (e) Presumed sperms between an oocyte and lyrate organ. Bar = 5 pm. (Q Detailed structure of a presumed sperm of (e). The detailed structure in the sperm cell is different from that shown in Di Palma Alberti (2001). Black bars are not a staining artifact but an unknown structure. Bar = 1 pm. Abbreviations Nl, N2, nuclei O, oocyte S, presmned sperm Yl, tipid-yolk granule Yp, protein-yolk granule (unpubHshed micrographs by Toyoshima Alberti).

Porter, K. R., Puck, T. T., Hsie, A. W., and Kelley, D., 1974, An electron microscopy study of the effects of dibutyryl cyclic AMP on Chinese hamster ovary cells. Cell 2 145. 

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