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Nikiyan H.N., Tatlybaeva E.B. ATOMIC FORCE MICROSCOPYUSING IN IDENTIFICATION OF SPECIFICALLY MARKED MOLECULES AND MICROORGANISM CELLSIdentification of biological molecules and microorganisms is an important problem of the microbiology which is traditionally solved by means of the marked antibodies. However in most cases the corresponding methods are based on detection of a significant amount of the interacting molecules and don't allow to find out their distribution on a surface. One of possible solutions consists in use of a modern method of atomic force microscopy. In current work the specific complexes formedbynanodimensionallabels, containing colloidal gold or amorphous carbon on a surface of bacterial cages of S. aureus and on a surface of plastic microwell immunological plateswere found and studied by means of atomic force microscopy. Morphological characteristics and the map of distribution of the revealed complexes on the studied surfaces were established. In experiments with tags sensitivity of a method was defined and comparison with the enzyme-linked immunoassay was carried out. The received results indicate high sensitivity of the offered method and certain advantages of carbon labels in comparison with gold labels, because of their easy detection and unambiguity in identification on the received images. Identification of microorganisms in model two-component biological systems was carried out and approaches to marking of bacterial cages in complex biological mixes was developed for the directed their allocation and the subsequent studying. Thus, use of a atomic force microscope in combination with nanodimensional carbon and gold labels, allows to realize a new way of identification of single molecules and microorganisms in complex biological mixes.Key words: molecules identification, microorganisms detection, gold and carbon labels, atomic force microscopy.
References:
1. Ramos-Vara J.A. Technical aspects of immunohistochemistry // Vet. Pathol. — 2005. — 42. — P. 405-426.
2. Electron Microscopy: Methods and Protocols, Kuo J ed. Humana Press. — 2007. — 608 p.
3. Binning G, Quate C.F, Gerber C. Atomic force microscope//Phys.Rev.Lett. — 1986. — 56(9). — P. 930-933.
4. Dufrкne Y.F., Hinterdorfer P. Recent progress in AFM molecular recognition studies//Pflugers Arch — Eur.J.Physiol. — 2008. — 256. — P.237–245 DOI:10.1007/s00424-007-0413-1.
5. Heinisch J.J, Lipke P.N, Beaussart A, El Kirat Chatel S, Dupres V, Alsteens D, Dufrкne Y.F.//J.Cell Sci. — 2012. — 125. — P. 4189-4195. — DOI: 10.1242/jcs.106005.
6. Maluchenko N.V., Agapov I.I., Tonevitsky A.G. et al. Detection of immune complexes using atomic force microscopy//Biofizika. — 2004. — V.49. — № 6. — Р. 1008–1014.
7. Quist, A. P. Direct measurement of single immunocomplex formation by atomic force microscopy/ A. P. Quist, A. A. Bergman, C. T. Reimann // Solution for a nanoscale world. — 2004. — № 8. — P. 4.
8. Nikiyan H., Tatlybaeva E., Rayev M. and Deryabin D. Applying Nanosized Gold and Carbon Immunolabels for the Quantitative Detection of Specific Ag–Ab Complexes by Using Atomic Force Microscopy//Current Nanoscience. — 2015. — V.11. — P. 615-620.
9. Tatlybaeva E.B., Nikiyan H.N., Vasilchenko A.S. and Deryabin D.G. Atomic force microscopy recognition of protein A on Staphylococcus aureus cell surfaces by labeling with IgG–Au conjugates//Beilstein J. Nanotechnol. — 2013. — 4. — P.743–749. DOI:10.3762/bjnano.4.84.
10. Edlich, R.F., Winters, K.L., Long, W.B., Gubler, K.D. Rubella and congenital rubella (German measles)// Journal of Long-Term Effects of Medical Implants. — 2005. — 15(3). — P.319–328.
11. Zhang, P.C., Bai, C., Ho, P.K., Dai, Y., Wu, Y.S. Observing interactions between the IgG antigen and anti-IgG antibody with AFM // IEEE Eng. Med. Biol. Mag. — 1997. — 16(2). — P.42-46.
12. Chen Y., Cai J., Xu Q., Chen Z.W. Atomic force bio-analytics of polymerization and aggregation of phycoerythrin-conjugated immunoglobulin G molecules//Mol. Immunol. — 2004. — 41(12). — P. 1247–1252. DOI 10.1016/ j.molimm.2004.05.012.
13. Forsgren A, Sjцquist J. "Protein A" from S. aureus. I. Pseudo-immune reaction with human gamma-globulin// J.Immunol. — 1966. — 97(6). — P.822–827.
14. DeDent A.C., McAdow M. and Schneewind O. Distribution of Protein A on the Surface of Staphylococcus aureus//J.Bacteriol. — 2007. — V.189. — №12. — P. 4473-4484.
15. Dickson J. S., Koohmaraie M. K. Cell surface charge characteristics and their relationship to bacterial attachment to meat surfaces //Appl.Environ.Microbiol. — 1989. — 55(4). — P.832.
16. Leff D.V., Brandt L., Heath J.R., Synthesis and Characterization of Hydrophobic, Organically-Soluble Gold Nanocrystals Functionalized with Primary Amines //Langmuir. — 1996. — 12 (20). — P. 4723–4730. — DOI 10.1021/la960445u.
About this article
Authors: Nikiyan A.N., Tatlybaeva E.B.
Year: 2015
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Editor-in-chief |
Sergey Aleksandrovich MIROSHNIKOV |
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