A protocol for Enhanced imaging and Quantification of Cervical Cell Under Scanning electron Microscope

Abstract

The application of Field Emission Scanning Electron Microscopy and Energy Dispersive X-Ray (FE-SEM/EDX) for the characterization of biological samples can produce promising results for classification purpose. The limitations of the established sample preparation technique of cervical cells for FE-SEM/EDX study that differentiate between normal and abnormal cells prompted the development of a proposed protocol for the preparation of cervical cells. The proposed protocol was conducted by a McDowell-Trump fixative prepared in 0.1M phosphate buffer without osmium tetroxide at 4°C for 2 h in the fixation process. Morphologically, the cervical cells scanned under the FE-SEM/EDX did not present blackening effects, and the structure of the cells was not broken based on the FE-SEM images. Quantitatively, the possible elemental distributions in the cells, such as carbon, nitrogen, oxygen, and sodium, are detected in samples prepared by the proposed protocol. The analysed elements were validated using the Attenuated Total Reflection and Fourier Transform Infrared (ATR/FTIR) spectroscopy. Moreover, by avoiding osmium tetroxide fixation, the time required for sample preparation decreased significantly. This sample preparation protocol can be used for normal and abnormal cervical cells in achieving better results in terms of morphological, detected elemental distribution, and rapid in time.