Skip to main content
. 2004 Jan;111(1):91–99. doi: 10.1111/j.1365-2567.2003.01782.x

Figure 1.

Figure 1

Topography of immunoreactive surfactant protein A (SP-A) in premenopausal vagina. (a) SP-A identified by immunocytochemistry (ICC), using Vector Red as the chromogen. (b) Preimmune and secondary antibody controls for ICC for SP-A using alkaline phosphatase reporter, and bromo-chloro-indolyl-phospahate (BCIP) and nitroblue tetrazolium (NBT) as the substrate and chromogen, respectively. (c) Correlation of localization of immunoreactive SP-A with glycogen. (a) Panel 1: at low magnification, SP-A (red reaction product) is seen in two distinct layers: in the basal portion of the intermediate layer, composed of rounded, metabolically active cells; and in the superficial layer, composed of flattened dead cells with pyknotic nuclei. Note the absence of immunostaining in the more superficial cells of the intermediate layer. The arrow points to one of the papillae comprising a core of subepithelial cells [shown in panels 2 and 3 of (a) by the letter ‘p’] surrounded by basal and parabasal cells. (a) Panels 2 and 3 show panel 1 at successively higher magnifications: panel 2 highlights the virtual absence of immunostaining in the more superficial portion of the intermediate layer, while panel 3 highlights the contrast between the strongly immunostaining cells in the intermediate layer and the minimal or no immunostaining of cells in the basal layer. The arrow points to the layer of cells demarcating the epithelium from the subepithelial layer (lamina propria). (b) Panel 1: immunoreactive SP-A (blue/black reaction product of BCIP/NBT) in a section from which glycogen had been removed by pretreatment with α-amylase. Note the similarity in localization of immunoreactive SP-A in the deep intermediate layer with that in sections containing glycogen (a, panel 1). (b) Panels 2 and 3: the absence of immunostaining in preimmune serum (panel 2) and secondary antibody (panel 3) controls. (c) Panels 1, 2 and 3: comparison of the topography of immunoreactive SP-A with that of glycogen. (c) Panel 1: section immunoreacted for SP-A after digestion of glycogen with α-amylase (blue/black reaction product of BCIP/NBT. (c) Panels 2 and 3: glycogen, stained magenta with pararosanaline, using periodic acid-Schiff (PAS) as the chromogenic dye, is seen throughout the intermediate layer. (c) Panel 4: elimination of staining for glycogen with PAS in a section pretreated with α-amylase. The stain for glycogen using pararosaniline in the PAS reaction, unlike basic fuchsin dye, does not stain the amylase-resistant intercellular material (see the Discussion). Sections were counterstained with haematoxylin in panels 1, 2 and 3 of (a), and in panels 2, 3 and 4 of (c). Images were recorded with a Nikon DXM 1200 digital camera using Nomarski optics and Nikon ACT1, version 2, software.