Correlative Microscopy of Ultrathin Cryosections is a Powerful Tool for Placental Research

Abstract

In this report, we describe procedures for correlative fluorescence and electron microscopy in immunocytochemical studies on the human placenta. Ultrathin cryosections of placenta were used for detection of the distribution of antigens by immunofluorescence and subsequently by immunoelectron microscopy of the same ultrathin cryosection. This methodology has certain advantages over conventional immunohistochemistry and immunoelectron microscopy. The advantages are, most notably, that the same exact structures are examined by both imaging modalities. In addition, since the tissue is physically sectioned (50–100nm thickness), greater resolution for fluorescence can be obtained in the z-dimension than can be obtained by optical sectioning in confocal microscopy. This last point is of particular importance for discriminating between structures closely stacked in the z-dimension. In this report, we have determined the distribution of caveolin-1 in ultrathin cryosections of terminal villi of the human term placenta. We demonstrate that the use of ultrathin cryosections is a powerful approach for immunofluorescence and correlative microscopy for the in situ localization of antigens.