Ultra-rapid non-invasive clinical nano-diagnostic of inflammatory diseases

Abstract

Several types of pathologies induce modified inflammatory responses in the organism, accompanied by changes in the circulating immune cells, with significant late effects and impacts on tissues organs and whole organism. Despite the significant progress of various diagnostic tools addressing many pathological situations, they still require elaborated and time-consuming laboratory work and analysis protocols. OBJECTIVES : Within the present study a new rapid atomic force microscopy (AFM) method based on concomitant isolation and fixation of specific living blood cell-type combined with direct AFM assessment is proposed. AFM provides a complex data overview on cell morphology in combination with quantification of morphometric parameters. Thus, it making possible a fine differentiation between disease activated- and normal-living cells. METHODS AND RESULTS : Blood monocytes are specifically isolated from the whole blood sample using CD14 antibody-coated magnetic beads, then immobilized on freshly cleaved mica surface and studied by means of environmental-ScanAsyst-AFM method. The cells were isolated, immobilized and washed in one step. Overview AFM images convey information to morphometric parameters such as surface roughness, height step, volume and aspect ratio, while higher resolution images resolve subsurface morphological features of human blood monocytes. CONCLUSIONS : A rapid method of isolation and selectively labeling the blood monocytes was developed, thus enabling a direct morphological assessment with AFM. Series of morphometric parameters such as surface roughness, height step, volume and aspect ratio are assigned and studied as potential diagnosis parameters. Multiplatform parameters noticeably help to differentiate monocytes present in different media as well as may be important for early diagnosis and later effective treatment. Such novel and rapid nano-diagnosis tools may constitute a “turning point” in prevention and disease detection from early stages before clinical symptoms appear, conveying towards valuable and indispensable clinical tools.