Existing users Log In New users Sign up

Non-invasive clinical nano-diagnostic

DISCOVERIES REPORTS (ISSN 2393249X), 2014, September-December issue


Rusu M, Dulebo A, Curaj A, Liehn EA. Ultra-rapid non-invasive clinical nano-diagnostic of inflammatory diseases. Discoveries Reports 2014, Sep-Dec; 1(1): e2. DOI: 10.15190/drep.2014.2

 Submitted: March 2nd, 2014; Revised: May 21st, 2014; Accepted: May 27th, 2014Published: May 28th, 2014;

 GO BACK to 2014, September-December issue


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

Mihaela Rusu (1), Alexander Dulebo (4), Adelina Curaj (1,2,3), Elisa A. Liehn (1,*)

(1) Institute for Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Germany
(2) Department of Thoracic, Cardiac and Vascular Surgery, RWTH Aachen University, Germany 
(3) Victor Babes National Institute of Pathology, Bucharest, Romania
(4) Bruker Nano Surfaces Division, Karlsruhe, Germany

*Correspondence to: Elisa A. Liehn, MD, PhD, Institute for Molecular Cardiovascular Research, University Hospital Aachen, Germany; Phone: +49-241-80 35983; Fax: +49-241-80 82716; Email: eliehn@ukaachen.de


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 sub-surface 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.


Access full text of the manuscript here: 


1. De Palma M, Murdoch C, Venneri MA, et al. Tie2-expressing monocytes: regulation of tumor angiogenesis and therapeutic implications. Trends in immunology. 2007;28:519-524.

2. Nahrendorf M, Swirski FK, Aikawa E, et al. The healing myocardium sequentially mobilizes two monocyte subsets with divergent and complementary functions. The Journal of experimental medicine. 2007;204:3037-3047.

3. Weber C, Zernecke A, Libby P. The multifaceted contributions of leukocyte subsets to atherosclerosis: lessons from mouse models. Nature reviews. 2008;8:802-815.

4. Lin SL, Castano AP, Nowlin BT, et al. Bone marrow Ly6Chigh monocytes are selectively recruited to injured kidney and differentiate into functionally distinct populations. J Immunol. 2009;183:6733-6743.

5. Ulrich C, Heine GH, Seibert E, et al. Circulating monocyte subpopulations with high expression of angiotensin-converting enzyme predict mortality in patients with end-stage renal disease. Nephrol Dial Transplant. 2010;25:2265-2272.

6. Karlmark KR, Weiskirchen R, Zimmermann HW, et al. Hepatic recruitment of the inflammatory Gr1+ monocyte subset upon liver injury promotes hepatic fibrosis. Hepatology 2009;50:261-274.

7. Zimmermann HW, Seidler S, Nattermann J, et al. Functional contribution of elevated circulating and hepatic non-classical CD14CD16 monocytes to inflammation and human liver fibrosis. PloS one. 2010;5:e11049.

8. Liehn EA, Postea O, Curaj A, et al. Repair after myocardial infarction, between fantasy and reality: the role of chemokines. Journal of the American College of Cardiology. 2011;58:2357-2362.

9. Liehn EA, Zernecke A, Postea O, et al. Chemokines: inflammatory mediators of atherosclerosis. Archives of physiology and biochemistry. 2006;112:229-238.

10. Subimerb C, Pinlaor S, Lulitanond V, et al. Circulating CD14(+) CD16(+) monocyte levels predict tissue invasive character of cholangiocarcinoma. Clinical and experimental immunology. 2010;161:471-479.

11. Kanzler I, Liehn EA, Koenen RR, et al. Anti-inflammatory therapeutic approaches to reduce acute atherosclerotic complications. Current pharmaceutical biotechnology. 2012;13:37-45.

12. Palmowski M, Huppert J, Ladewig G, et al. Molecular profiling of angiogenesis with targeted ultrasound imaging: early assessment of antiangiogenic therapy effects. Molecular cancer therapeutics. 2008;7:101-109.

13. Palmowski M, Morgenstern B, Hauff P, et al. Pharmacodynamics of streptavidin-coated cyanoacrylate microbubbles designed for molecular ultrasound imaging. Investigative radiology. 2008;43:162-169.

14. Fokong S, Fragoso A, Rix A, et al. Ultrasound molecular imaging of E-selectin in tumor vessels using poly n-butyl cyanoacrylate microbubbles covalently coupled to a short targeting peptide. Investigative radiology. 2013;48:843-850.

15. Wu Z, Curaj A, Fokong S, et al. Rhodamine-Loaded Intercellular Adhesion Molecule-1-targeted Microbubbles for Dual-Modality Imaging Under Controlled Shear Stresses. Circ Cardiovasc Imaging. 2013;6:974-981.

16. Orbay H, Zhang Y, Valdovinos HF, et al. Positron emission tomography imaging of CD105 expression in a rat myocardial infarction model with (64)Cu-NOTA-TRC105. American journal of nuclear medicine and molecular imaging. 2013;4:1-9.

17. Buonincontri G, Methner C, Carpenter TA, et al. MRI and PET in Mouse Models of Myocardial Infarction. J Vis Exp. 2013.

18. Gargiulo S, Greco A, Gramanzini M, et al. PET/CT imaging in mouse models of myocardial ischemia. Journal of biomedicine & biotechnology. 2012;2012:541872.

19. Lim TS, Ricciardi-Castagnoli P. Single-cell force spectroscopy: mechanical insights into the functional impacts of interactions between antigen-presenting cells and T cells. Immunologic research. 2012;53:108-114.

20. Starodubtseva MN, Yegorenkov NI, Nikitina IA. Thermo-mechanical properties of the cell surface assessed by atomic force microscopy. Micron. 2012;43:1232-1238.

21. Wang Z, Liu L, Wang Y, et al. A fully automated system for measuring cellular mechanical properties. Journal of laboratory automation. 2012;17:443-448.

22. Reichlin T, Wild A, Durrenberger M, et al. Investigating native coronary artery endothelium in situ and in cell culture by scanning force microscopy. Journal of structural biology. 2005;152:52-63.

23. Lim TS, Mortellaro A, Lim CT, et al. Mechanical interactions between dendritic cells and T cells correlate with T cell responsiveness. J Immunol. 2011;187:258-265.

24. Marsh G, Waugh RE. Quantifying the mechanical properties of the endothelial glycocalyx with atomic force microscopy. J Vis Exp. 2013:e50163.

25. Natkanski E, Lee WY, Mistry B, et al. B cells use mechanical energy to discriminate antigen affinities. Science 2013;340:1587-1590.

26. Saliba AE, Saias L, Psychari E, et al. Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays. Proceedings of the National Academy of Sciences of the United States of America. 2010;107:14524-14529.

27. Skorkina MY, Fedorova MZ, Muravyov AV, et al. The use of nanomechanic sensor for studies of morphofunctional properties of lymphocytes from healthy donors and patients with chronic lymphoblastic leukemia. Bulletin of experimental biology and medicine. 2012;154:163-166.

28. Braga PC, Ricci D. Atomic Force Microscopy. Biomedical Methods and Applications. Totowa, New Jersey: Humana Press; 2004: 394.

29. Dulebo A, Preiner J, Kienberger F, et al. Second harmonic atomic force microscopy imaging of live and fixed mammalian cells. Ultramicroscopy. 2009;109:1056-1060.

30. Espenel C, Giocondi MC, Seantier B, et al. Temperature-dependent imaging of living cells by AFM. Ultramicroscopy. 2008;108:1174-1180.

31. Pandiyaraj KN, Deshmukh RR, Mahendiran R, et al. Influence of operating parameters on surface properties of RF glow discharge oxygen plasma treated TiO2/PET film for biomedical application. Materials science & engineering. 2013;36:309-319.

32. Alsteens D, Dupres V, Mc Evoy K, et al. Structure, cell wall elasticity and polysaccharide properties of living yeast cells, as probed by AFM. Nanotechnology. 2008;19:384005.

33. Fuhrmann A, Staunton JR, Nandakumar V, et al. AFM stiffness nanotomography of normal, metaplastic and dysplastic human esophageal cells. Physical biology. 2011;8:015007.

34. Kong F, Li Z, Parks WM, et al. Cyclic mechanical reinforcement of integrin-ligand interactions. Molecular cell. 2013;49:1060-1068.

35. El Kirat K, Burton I, Dupres V, et al. Sample preparation procedures for biological atomic force microscopy. Journal of microscopy. 2005;218:199-207.

36. Francis LW, Gonzalez D, Ryder T, et al. Optimized sample preparation for high-resolution AFM characterization of fixed human cells. Journal of microscopy. 2010;240:111-121.

37. Fojt L, Klapetek P, Strasak L, et al. 50 Hz magnetic field effect on the morphology of bacteria. Micron. 2009;40:918-922.

38. Chen D, Gan H, Huang X, et al. Effects of peripheral blood mononuclear cells morphology on vascular calcification in uremic patients on maintenance hemodialysis. Ther Apher Dial. 2012;16:173-180.

39. Tsirpanlis G, Chatzipanagiotou S, Ioannidis A, et al. Serum and peripheral blood mononuclear cells infectious burden: correlation to inflammation and atherosclerosis in haemodialysis patients. Nephrology 2005;10:256-263.

40. Leabu M. The still valid fluid mosaic model for molecular organization of biomembranes: accumulating data confirm it. Discoveries. 2013;1:e7.

41. Nicolson GL. Update of the 1972 Singer-Nicolson Fluid—Mosaic Model of Membrane Structure. Discoveries. 2013;1:e3.

42. Zhang W, Gao Z, Shao D, et al. Atomic force microscopy analysis of progenitor corneal epithelial cells fractionated by a rapid centrifugation isolation technique. PloS one. 2013;8:e59282.

43. Song J, Xiao LH, Hei Y, et al. Study on peripheral blood T lymphocyte cell membrane in thyroid associated ophthalmopathy by using atomic force microscopy. Chinese journal of ophthalmology. 2009;45:522-527.

44. Kim KS, Cho CH, Park EK, et al. AFM-detected apoptotic changes in morphology and biophysical property caused by paclitaxel in Ishikawa and HeLa cells. PloS one. 2012;7:e30066.

45. Kliche K, Kuhn M, Hillebrand U, et al. Direct aldosterone action on mouse cardiomyocytes detected with atomic force microscopy. Cell Physiol Biochem. 2006;18:265-274.

46. Oberleithner H, Peters W, Kusche-Vihrog K, et al. Salt overload damages the glycocalyx sodium barrier of vascular endothelium. Pflugers Arch. 2011;462:519-528.


News & Events Latest news from Discoveries Reports

  • 2015 | Indexed by Google Scholar

    All our published articles are now indexed by Google Scholar! First citations to articles published in Discoveries Reports are included! Search for the article's title (recommended) or the authors:

    Google Scholar Search
  • 2014 | Discoveries Reports

    DOIs (Digital Object Identifiers) are now assigned to all our published manuscripts in Discoveries Reports. DOI uniquely identifies an article and is provided by CrossRef.

  • 2014 | Manuscript Submission

    Submit your manuscript FREE, FAST and EASY ! (in less than 1 minute)
    There are NO fees for the manucript submission or publishing of the accepted manuscripts.

    read more

    We are now ACCEPTING MANUSCRIPTS for DISCOVERIES REPORTS, publishing inovative and important research findings from all areas related to Medicine, Biology and Chemistry. We are also accepting experimental articles that validate/invalidate highly used reagents in current publications (ex. antibodies) and selected articles presenting negative data with impact and of wide scientific interest ...

    read more
Member Login
Free Registration Click here to sign up
Copyright © 2013 Applied Systems. All Rights Reserved.