Useing AFM To Observe The Cytomembrane Surface Morphology Of Human Oral Squamous Epithelium Combine With Membrane Antibodyes

ObjectiveUsing Atomic Force Microscope(AFM) to observe cytomembrane surfacemorphology of human oral squamous epithelial cells before and after combiningwith membrane antibodies.Providing morphology figure of antibody binding tothe surface of cells on nanometer level,which provides a new method forimmunohistochemistry nanometer level research, antigen accurate location anddeeply understanding of exception expressed phenomenon.MethodsClients are divided into three groups according to different reagents:PBScontrol group, EMA antibody experimental group and S-100antibodyexperimental group. Inclusion criteria:1.From twenty to thirty-five yearsold,male;2.No oral diseases;3.No local affective factors;4.No experience ofbeing in touch with radiations and poison;5.No smoking;6.Agree withcooperation. Using the method of cells smear and selecting the AFM contactmode to scan cytomembrane surface structure of normal human oral squamousepithelial cells. Applicating the AFM image analysis software to measure andanalysis the important cell membrane parameters of Mean Roughness (Ra) andThe Maximum Height (Rmax).Then All data were analyzed by using SPSS13.0 statistical software.Results1pathological cytology resultsThe results of each group of human oral squamous epithelial cells withimmunohistochemical stains in light-microscope have been observed by twopathological diagnosis doctors.Cells in EMA antibody experimental group showpositive reactions on cytomembrane while negative reactions on cytoplasm andcyteblast.Cells in PBS control group and S-100antibody experimental groupshow negative reactions on cytomembrane,cytoplasm and cyteblast.Howeverthere are no significant changes in morphology of the cyomembrane.2AFM cell scanAFM was applicated to observe oral squamous epithelial cells of eachgroup.①．Adding PBS,EMA antibody,S-100antibody respectively and begin toscan.The cells contour of each group have no significant differences. Cells aremostly flat, irregularly appearance like round,oval or polygon.②We found thatthe surface of the control group cells is ups and downs.The upheavals arrangeirregularly like hellolls or mountains.There forms pits between upheavals withdifferent width and depth. Compared with the control group,the cytomembranesurface form of EMA antibody experimental group shows that there are somegranular materials adhere to the cytomembrane surface.The granular materialsare similar size, regular shape, any number, uneven distribution. By partialscaning we could obseve that the granular materials show us a peak shapestructure which attach to the “helloll”or”mountain” surface.The average width ofthe peak shape structure is about2μm,and the average height is about600nm.The upheavals of the cytomembrane surface are more higher,the ups and downs and roughness are more obvious. There are no obvious differencesin the mophology of sytomembrane surface between S-100antibodyexperimental groups and control group.3Statistical analysis resultsAll data were divided into three groups: control group.We analyzed all Raand Rmax values of each image and used SPSS13.0software for processing.All experimental data were indicated in form ofx±s.The Ra values(nm) of eachgroup are32.799±1.769，61.168±2.863，33.317±1.845.The Rmax values(nm)of each group are253.33±1.34，572.863±2.363，248.87±1.587. There arestatistical differences between the EMA antibody experimental group and thecontrol group in Ra and Rmax values of Oral squamous epithelial cell surface（p<0.05）. There is no significantly difference between the S-100antibodyexperimental group and the control group in Ra and Rmax values of oralsquamous epithelial cell surface（p>0.05）.conclusionUsing atomic force microscope to observe the cytomembranesurface morphology of human oral squamous epithelia cell binded to membraneantibodies shows that Most of the cells have granular substanceattached.Height is about600nm,width’s about2μm, any number,unevendistribution,Ra and Rmax are both increased compared with the controlgroup.There is no obvious change on the cells that without membrane antibodycombine with. The specific change of cytomembrane surface binded toantibodys has provides a new thinking of clinical diagnosis on nanoscale level,and lays the foundation for the following immunohistochemical nanoscalestudy.AFM might become a valuable auxiliary diagnostic tool in clinical diagnosis.