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Single-Molecule Measurements in Solution by FCS/FCCS:

Abstract: FWF Austrian Science Fund Project P20454-N13: Stand-Alone Collaborative Research Project with visiting professorships (Forschungsprofessuren) in the USA, Japan and Cairo for about 27 months: dienstzuteilung aboard (im Ausland).

The work presented in this authorized and supported project program involves the development of a new analysis technique for single-molecule spectroscopy and imaging. For the first time, it is turned into a potential opportunity. The new concept of analysis of just one single molecule in solution over extended periods of time will make it possible to observe the single molecules without bias to the millisecond range in solution of freely diffusing molecules or to artificial immobilization on surfaces or hydrodynamic focusing. The following main idea is put forward: is it possible to statistically recognize the same molecule multiple times as it diffuses through a small confocal probe volume? If so, the detection time of an individual molecule freely diffusing in solution could be extended. For the new analysis approach, it is crucial to understand the balance between the time scale and the frequency that a single molecule reenters the confocal probe volume and the overall time necessary for the measurement.
The proposed research will provide with a better understanding of some of the theoretical and experimental backgrounds of these types of analyses and biological or medical applications. The originality and innovative nature of the proposal work reside in the strict coupling of latest microscopic and spectroscopic developments with physiological relevance. The new ideas are comprehensively presented and this relationship is a new concept at the time. Possible users for this concept are those working in biotechnological applications dealing with gene technology. Furthermore, the concept is of interest for a great number of medical, pharmaceutical and bio-chemical laboratories. It may serve as a foundation for further work in single-cell biology and immunology.

URL: http://topics.scirus.com/Measurements_of_Single_Molecules_in_Solution_and_Live_Cell_at_Time_Scales_That_Are_Currently_Not_Available_Impact_on_Immunology.html

Keywords: applied mathematics; numerical analysis; biophysics; optics; analytical chemistry; biochemistry; physical chemistry; biotechnology; medical biochemistry; medical molecular biology; Immunology; Laser gun; Molecular Medicine; New technique for single-molecule spectroscopy and imaging; Optical engineering

Local Subprojectlead:: Földes-Papp Zeno

Duration:: 01.12.2007-20.06.2011

Programme:: Einzelprojekt

Type of Research: basic research

Staff: Földes-Papp, Zeno, Project Leader

MUG Research Units: Department of Internal Medicine, Joint Facilities

Funded by: FWF, Fonds zur Förderung der Wissenschaftlichen Forschung, Wien, Austria

Project results published: > Fluorescence molecule counting for single-molecule... Curr Pharm Biotechnol. 2011; 12(5):824-833; > Single actomyosin motor interactions in skeletal m... Biochim Biophys Acta. 2011; 1813(5):858-866; > Meaningful interpretation of subdiffusive measurem... Curr Pharm Biotechnol. 2010; 11(5):527-543; > Defining features and exploring chemical modificat... Curr Pharm Biotechnol. 2010; 11(5):518-526; > Confocal fluctuation spectroscopy and imaging.... Curr Pharm Biotechnol. 2010; 11(6):639-653; > Long-term marker-free multiphoton imaging, targete... Invited paper. SPIE Conference Proceedings. Session 6: Technology Development and Applications I Date: Monday 25 January Time: 1:35 PM - 5:20 PM Room: Room 308 (Esplanade)2010; -SPIE Photonics West; JAN 23-28, 2010; San Francisco, CA, USA.; > Nanoprocessing and targeted transfection of stem c... Invited paper. SPIE Conference Proceedings. Session 1: Cell Imaging Date: Saturday 23 January Time: 8:30 AM - 12:00 PM Room: Room 200 (Mezzanine)2010; -SPIE Photonics West; JAN 23-28, 2010; San Francisco, CA, USA.; > Anomalous behavior in length distributions of 3D r... Opt Express. 2010; 18(17):17883-17896; > Long-term marker-free multiphoton imaging, targete... SPIE, Vol. 7569, 756916-756916-7 ; doi:10.1117/12.842024 . 2010; > Myocardial expression of TNF-alpha, IL-1beta, IL-6... Curr Pharm Biotechnol. 2010; 11(5):413-420; > Polarized fluorescent nanospheres.... Opt Express. 2010; 18(5):4289-4299; > NIH site-visit preparation - single molecules. ZFP... 8th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; MAR 28-APR 05, 2009 ; Dallas-Fort Worth, Texas, USA. 2009.; > Real-Time Observation of Individual Particles and ... http://www.aspenphys.org/ ; Proceedings of the Research Conference on "Single Molecule Biophysics" 2009; -Research Conference on "Single Molecules" - ASPEN Center of Physics. Organized by Departments of Biological Sciences and Applied Physics, Stanford University, Stanford, CA. ; JAN 4-10, 2009; Aspen, Colorado, USA.; > Fluorescence Microscopy and Spectroscopy I. ZFP ac... 1st Working Stay at the Helen Diller Family Comprehensive Cancer Research Center of the University of California San Francisco.; JUNE 07-20, 2009; University of California San Francisco, CA, USA. 2009.; > Comparative study of the immunological marker IL-6... The Open Biochemistry Journal. 2009; 3: 124-130.; > Measurements of cytomegalovirus fluorescence fluct... 10th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; JUL 25 - AUG 02, 2009; Dallas-Fort Worth, Texas, USA. 2009.; > Simulations of Single-Molecule Measurements in Sol... 7th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; JAN 11-26, 2009 ; Dallas-Fort Worth, Texas, USA. 2009.; > Reducing background contributions in fluorescence ... Curr Pharm Biotechnol. 2009; 10(5):532-542; > Chapter 48: Viral Chip Technology in Genomic Medic... In: Willard, HF; Ginsburg, GS editors(s). Handbook of Genomic and Personalized Medicine. 2009. [Publication of the National Center for Human Genome Sequencing and Personalized Medicine of the USA] . vol. 1: New York: Elsevier Inc.; 2009. p. 538-561. (ISBN: 0-12-369420-5); > Chapter 13: Confocal microscopy (original paper). ... In: Doucette R; Walz, W; editors(s). Imaging and Neurohistology: Basic Techniques. Upcoming in 2009, in press. Totowa, NJ, USA: Humana Press, Inc.; 2009.; > Trafficking of mature miRNA-122 into the nucleus o... Curr Pharm Biotechnol. 2009; 10(6): 569-578.; > Experimental fluorescence fluctuation time traces.... 9th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; JUL 11-20, 2009; Dallas-Fort Worth, Texas, USA. 2009.; > Fluorescence Microscopy and Spectroscopy II. ZFP a... 2nd Working Stay at the Helen Diller Family Comprehensive Cancer Research Center of the University of California San Francisco.; AUG 03 - AUG 15, 2009; University of California San Francisco, CA, USA. 2009.; > Single-Molecule Fluorescence Correlation Spectrosc... Invited Seminar in the Health Science Center of the University of North Texas, Department of Molecular Biology and Immunolgy; MAR 12-14, 2008; Dallas (Texas), USA. 2008.; > Measurements of Single Molecules. part III. ZFP ac... 4th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; AUG 22 - SEP 13; Dallas-Fort Worth, Texas, USA. 2008.; > Single-Molecule FCS and FCCS in Solution.... Proceedings of the 7th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine and the 11th International Carl Zeiss-sponsored Workshop on Fluorescence Correlation Spectroscopy (FCS) and related Methods held as part of the Weber Symposium: http://www.lfd.uci.edu/weber/symposium/ 2008; -The 7th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine; JUN 6-12, 2008; Kauai (HAWAII), USA. (ISBN: http: ); > Invited Lecture: Single-Molecule Fluorescence Corr... Seminar in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; MAR 12-14, 2008; Dallas-Fort Worth, Texas, USA. 2008.; > Measurements of Single Molecules. part IV. ZFP ack... 5th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center and Annual Retraeat of the Department of Molecular Biology and Immunology at the University of Oklahoma Biological Station at Lake Texoma from Oct 16-18, 2008; OCT 07-21; Dallas-Fort Worth, Texas, USA. 2008.; > Measurements of a Single-Molecule at Very Low Conc... 2nd Five Days' Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; APR 27 - MAY 03, 2008; Dallas-Fort Worth, Texas, USA. 2008.; > Measurements of a Single-Molecule at Very Low Conc... 3rd Five Days' Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center; MAY 18 - MAY 24, 2008; Dallas-Fort Worth, Texas, USA. 2008.; > Measurements of Single Virus particles. ZFP acknow... 6th Working Stay in the Department of Molecular Biology and Immunology, University of North Texas Health Science Center ; DEC 08-14, 2008 ; Dallas-Fort Worth, Texas, USA. 2008.; > What is the meaningful time for studying a single ... Proceedings of the 7th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine and the 11th International Carl Zeiss-sponsored Workshop on Fluorescence Correlation Spectroscopy (FCS) and related Methods held as part of the Weber Symposium2008; -7th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine: http://www.lfd.uci.edu/weber/symposium/; JUN 6-12, 2008; Kauai, Hawaii, USA.; > Exploring the biomedical applications of microscop... EXP MOL PATHOL. 2007; 82(2): 103-103.; > Chapter 5.6.6 General considerations of crosstalk ... In: Carl Zeiss Advanced Imaging Microscopy; editors(s). Applications Handbook LSM 510/Confocor: Fluorescence Correlation Spectroscopy, B40-641e 05/01 version 7, Zeiss, Germany. 2007.; > Invited Speaker: Theoretical testing of single-mol... Proceedings of the 10th International Workshop on Fluorescence Correlation Spectroscopy and Related Methods. http://www.lfsci.hokudai.ac.jp/FCS-2007/10th/sub4.html ,2007; -10th International Workshop on Fluorescence Correlation Spectroscopy and Related Methods, (Z. Földes-Papp is sponsored by the Hokkaido University, Graduate School of Life Science, Laboratory of Molecular Cell Dynamics , Sapporo, Japan); November 26-28, 2007; Sapporo, JAPAN .; > Mobility and distribution of replication protein A... Exp Mol Pathol. 2007; 82(2):156-162; > Fluorescence fluctuation spectroscopic approaches ... Curr Pharm Biotechnol. 2007; 8(5):261-273; > "True" single-molecule molecule observations by fl... Exp Mol Pathol. 2007; 82(2):147-155; > What it means to measure a single molecule in a so... Exp Mol Pathol. 2006; 80(3):209-218; > Keynote Speaker: True single-molecule molecule m... Proceedings of the 30th Annual Meeting of the Microscopical Society of Ireland2006; -30th Annual Meeting of the Microscopical Society of Ireland, (Z. Földes-Papp is sponsored by the National University of Ireland); http://www.nuigalway.ie/msi/keynote2006.htm ; August 30-September 1, 2006; Galway, IRELAND.