Zastosowanie Wzmocnionej Powierzchniowo Spektroskopii Ramana (SERS) w Detekcji Materiałów Biologicznych

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Bogdan Marszałek
10 lat temu
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1 Zastosowanie Wzmocnionej Powierzchniowo Spektroskopii Ramana (SERS) w Detekcji Materiałów Biologicznych Bartosz BARTOSEWICZ, a Magdalena GAJDA-RĄCZKA, a Małgorzata SZPAKOWSKA, b Elżbieta TRAFNY, b Bartłomiej J. JANKIEWICZ a a Institute of Optoelectronics, Military University of Technology, 2 Kaliskiego Str., Warsaw b Military Institute of Hygiene and Epidemiology, 4 Kozielska Str., Warsaw

2 Zagrożenie Atakiem Terrorystycznym Biosecurity Effort Expands To Africa - Chemical & Engineering News 2011 The Cooperative Threat Reduction (CTR) program - U.S. effort to secure deadly pathogens to prevent their use in bioterror attacks. Some of the world s deadliest diseases including the Ebola, Marburg, and Rift Valley Fever viruses occur naturally in Africa, a volatile region of the world where civil upheaval and terrorism are widespread. A Ugandan facility that Senator Lugar visited lacks the proper tools to handle samples carrying deadly diseases such as anthrax and Ebola virus. A handwritten logbook at a lab in Uganda notes a suspected case of ANTHRAX outbreak. Chemical & Engineering News Copyright 2013 American Chemical Society

Some of the world s deadliest diseases including the Ebola, Marburg, and Rift Valley Fever viruses occur naturally in Africa, a volatile region of the world where civil upheaval and

3 Detekcja i identyfikacja materiałów biologicznych Polymerase Chain Reaction (PCR, molecular method) What about spectroscopic methods? immunological methods MALDI-TOF Spectrometry microscopic examination

4 Metody spektroskopowe badania materiałów biologicznych FT-IR spectroscopy Fluorescence excitation-emission maps of selected Fungi Raman Spectroscopy

5 E 0 (λ) absorption emission scattering molecule PROBLEM!!! Low signal intensity I I Ram Ray Spektroskopia Ramana 10 4 E 1 H. Barańska, A. Łabuzińska, J. Trepiński, Laserowa spektrometria ramanowska, zastosowania analityczne, Warszawa Rayleigh scattering hν 0 Raman scattering hν R ν 1 ν w ν 0

6 Fotowoltaika Materiały elektroniczne Kataliza Reaktywność : Zastosowanie Spektroskopii Ramana Krzem Półprzewodniki Badanie sieci krystalicznych Badanie przebiegu reakcji Polimorfizm Nanowęgiel Fizyka Chemia Farmacja SERS/TERS Nanotechnologia Raman i SERS Biologia Biodiagnostyka Inżynieria wytwarzania Inżynnieria materiałowa Geologia Obrazowanie tkanek i komórek Polimery Pokrycia powierzchni Badania minerałów Sądownictwo Żywność Ceramika Gemmologia Sztuka Identyfikacja materiałów

Nanowęgiel Fizyka Chemia Farmacja SERS/TERS Nanotechnologia Raman i SERS Biologia Biodiagnostyka Inżynieria wytwarzania

7 Wzmocniona Powierzchniowo Spektroskopia Ramana N. Chandrabhas, S. Soumik, Nanomater. Nanotechnol. 2012, 2, 1-13,

8 Nanostruktury plazmonowe Plazmonika jest obszarem nanofotoniki związanym głównie z manipulowaniem światła w nanoskali w oparciu o właściwości plazmonów powierzchniowych, czyli oscylacji elektronów swobodnych w strukturze metalicznej wzbudzonych przez falę elektromagnetyczną. Nanostruktury plazmonowe czyli nanostruktury charakteryzujące się występowaniem plazmonów powierzchniowych obejmują między innymi warstwy metaliczne, jak również nanocząstki metaliczne. 500nm

metalicznej wzbudzonych przez falę elektromagnetyczną.

9 Zastosowanie nanostruktur plazmonowych

10 Nanoshell Terapia Nowotworowa >90% remission achieved in mice, rats, rabbit animal models in independent sites and studies No toxic effects observed, even when dossage >20X projecthuman dose NS accumulate in liver and are slowly removed over several weeks (feces) Projected human dose: ~1.4 a per 70 kg patient for untargeted therapy (approximately $30 of Au) patients treated per oz of Au NS photothermal therapy approved January 2008 for clinical trials Halas et al. PNAS 2003;100: ; Cancer Letters 209 (2004)

11 Nanoshell wzmocnienie sygnału w SERS i SEIRA H.Wang, J. Kundu, N.J. Halas, Angew. Chem. Int. Ed., 46, 9040 (2007) SERS and SEIRA spectra of pma Enhancement: SERS SEIRA 10 4

12 SERS Bakterii Przegląd literaturowy It was shown that use of PCA and SERS spectra allows to discriminate between different species in this case different strains of Bacillus spores. In addition, studies have shown that Gram-positive Bacillus spores can be differentiated from Gram-negative vegatative cells. Guicheteau et al. Appl. Spectrosc. 2008, 62, 267

13 SERS Bakterii Przegląd literaturowy The multivariate data analysis of the SERS spectra of bacteria prepared by the standard multiple washing/centrifugation protocol shows that different bacterial species grown in the same media exhibit different SERS spectra, the same bacterial species grown in different media show the same SERS spectra, and, finally, SERS spectra of bacteria and the culture media in which they are grown are reproducibly distinct. Ziegler et al. Appl. Spectrosc. 2011, 65, 493

SERS spectra, the same bacterial species grown in different media show the same SERS spectra, and, finally, SERS spectra of

14 SERS Bakterii Przegląd literaturowy A rapid detection protocol suitable for use by first-responders to detect anthrax spores using a low-cost, battery-powered, portable Raman spectrometer has been developed. Van Duyne reports that the procedure developed in his lab takes only 11 minutes and is sensitive enough to detect about 2,600 anthrax spores--about one-quarter the infectious dose of 10,000 spores.

15 SERS Bakterii Przegląd literaturowy Marotta et al. Appl. Spectrosc. 2010, 64, 601 Entities such as mammalian cells and spores give broader, more complex spectra. The origin of these signals is likely to be mainly from the part of the cell or spore closest to the enhancing surface and various bands that are indicative of proteins can be identified. Given the complex nature of the spectra, a reliable substrate and correct deposition techniques are again required. The data analysis by methods such as principal component analysis (PCA) allows to discriminate between genetically different species of intact bacillus spores. There are still problems to solve G. McNay et al. Appl. Spectrosc. 2011, 65, 825

identified. Given the complex nature of the spectra, a reliable substrate and correct deposition techniques are again required.

16 Wytwarzanie nanostruktur plazmonowych w IOE WAT

17 SERS Bakterii badania w IOE WAT Surface Enhanced Raman Spectroscopy (SERS) application of surface plasmon resonance of Au and Ag nanoparticles to enhance Raman scattering signal of analyte. Au Intensywność (a.u.) Liczba falowa (cm -1 ) Ag SERS spectrum of single Bacillus thuringiensis cell on Ag colloids

18 SERS Bakterii badania w IOE WAT 200 nm SERS spectrum of single Bacillus thuringiensis cell on core-shell nanostructures SiO

19 SERS Bakterii badania w IOE WAT SiO AgNPs(Turkevich) AgNPs(HCHO) AuNPs(Turkevich)

20 EDA Joint Investment Programme on CBRN Protection (JIP CBRN) RAMBO - Rapid Air-particle Monitoring against BiOlogical threats Project Duration: (36 months) Overall Proposal Cost SELEX-Sistemi Integrati (SSI) Consorzio CREO Centro Ricerche Elettro- Ottiche (CREO) ENEA microfluidic ChipShop IOE WAT Universite Claude Bernard Lyon (UCBL) IT IT IT DE PL FR Polish partners of IOE WAT in RAMBO project: Epidemiological Response Centre of Polish Armed Forces Military Institute of Hygiene and Epidemiology

000 SELEX-Sistemi Integrati (SSI) Consorzio CREO Centro Ricerche Elettro- Ottiche (CREO) ENEA microfluidic ChipShop IOE WAT

21 Project Description The RAMBO project aims at developing advanced methods, instrumentation and sensing strategies/protocols for continuous monitoring of air particles against biological threats and in particular the Anthrax (through an experiment on its stimulants). More specifically, RAMBO aims at developing an advanced sensor with the following performance objectives: Good selectivity (low probability of False Alarms) Detect to Warn response time (<45 min) Minimal use of chemicals and reagents (for long-term unattended operation) State of the art sensitivity (towards the one-spore limit) Man-Portability and ruggedizability for use in defence and security scenarios Broad biological spread (investigations could be later extended to virus and toxins in that same list) The objective of RAMBO is to investigate and apply the 2 sensing techniques to the detection and classification of bio agents (actually stimulants of real agents). The SERS (Surface Enhanced Raman Spectroscopy) cell functionalised with Phages long life specific receptors, will be connected through a microfluidic path to commercially available air samplers upstream and downstream to PCR (Polymerase Chain Reaction) for classification/identification of the bio agents. The B -point detection sensor will operate as a first alarm detector, and will also allow the identification of the threat. It will be implemented in an portable device that can be left unattended for long time.

22 Thank you for your attention!

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