Consecutive light microscopic, transmission and scanning electron microscopic study of the same exfoliated cells from human uterine cervix.

Exfoliative squamous cells on a slide-glass-sized x-ray film obtained from the uterine cervix, were examined by light microscopy (LM). The portion of the smear where the target cells were observed was cut out, dried by the critical point drying technique (CPD), and examined by scanning electron microscopy (SEM). Then the target cells were embedded in Epon 812 Resin. Ultrathin sections were made and examined by transmission electron microscopy (TEM). The preparation for this study was as follows.<BR>1) For TEM examination, the cells were fixed with paraformaldehyde, glutalaldehyde and osmic acid. But the fixation resulted in a marked change in the color of Papanicolaou staining. To avoid this discoloration, the Papanicolaou's method was modified.<BR>2) CPD, used for preparing the cells for SEM examination, mode the cells insufficient to the penetration of Epon 812 Resin, which was necessary to preserve the intracellular ultrastructure for the subsequent TEM examination. To resolve the problem, complete rehydration of the cells after SEM examination was performed. Thus, it became possible to make a study of cells through consecutive LM-SEM-TEM examination.<BR>Using the technique, cells of the superficial, intermediate and basal layers of the squamous epithelium, dyskaryotic cells of the intermediate layer, a dysplastic cell from a case of squamous cell carcinoma which was diagnosed by LM as a degenerated megalocyte, an atypical cell collected in swabs from a condylomatous lesion and atypical cells of the superficial layer in a Vira Pap positive case were examined by the consecutive method. As a result, many new findings which have never been observed were seen. A technique which allows the examination of the same exfoliated cells by the consecutive LM-SEM-TEM method has almost been established and the usefulness of this technique has been demonstrated.