Single-Molecule/Entity Spectroscopy is an indispensable resource for scientists interested in understanding single-molecule spectroscopy comprehensively. By presenting detailed information on the technique’s principles, discussing its diverse applications, and providing insights into future developments, researchers can acquire a holistic overview of this dynamic field without the need to consult an extensive array of individual papers. This primer bridges the gap between standard undergraduate coursework to practical implementation in the research lab for single-molecule spectroscopy.

Single Molecule Spectroscopy is one of the hottest topics in today's chemistry. It brings us close to the the most exciting vision generations of chemists have been dreaming of: To observe and examine single molecules! While most of chemistry deals with myriads of molecules, this books presents the latest developments for the detection and investigation of single entities. Written by internationally renowned authors, it is a thorough and comprehensive survey of current methods and their applications.

The topics range from single molecule experiments in quantum optics and solid-state physics to analogous investigations in physical chemistry and biophysics.

Written by the leading experts in the field, this book describes the development and current state of the art in single molecule spectroscopy. The application of this technique, which started 1989, in physics, chemistry and biosciences is displayed.

Analytical measurements at the single molecule level under ambient conditions have become almost routine in the past few years. The application of this technology to fundamental studies of heterogeneity in biomolecular structure and dynamics, chemical and biological reaction kinetics, and photophysics provides a rich playground for molecular scientists. The potential use of single molecule detection for nanotechnology and quantum information processing is a new and almost unexplored area. This handbook is intended for those interested in a practical introduction to single molecule investigations using fluorescence techniques and places special emphasis on the practicalities of achieving single molecule resolution, analysing the resulting data and exploration of the applications in biophysics. It is ideal for graduate research students and others embarking on work in this exciting field.

Single molecule fluorescence spectroscopy has attracted considerable attention over the past two decades. Measurement on a single entity provides an opportunity to avoid ensemble averaging which is always present in conventional bulk fluorescence measurements. This makes single molecule spectroscopy particularly interesting for biophysics and biochemistry where heterogeneous systems are often encountered. The general interest of this thesis is in studies of single immobilised molecules carried out at room temperature. One of t.he major issues of single molecule spectroscopy is finding a suitable immobilising medium. Inorganic silica matrices prepared by the sol-gel method have a great potential to provide a close- to-natural immobilising environment even for sensitive biomolecules and thus allow investigation of their natural behaviour on the most fundamental level. In order to be able to tailor both physical and chemical properties of the final gel, it is of great importance to develop reliable methods to control each stage of polymerisation. In one part of this thesis, applications of fluorescent probes to investigation of sol-gels properties, as well as monitoring the gel assembly process, are discussed. The thesis further presents studies of the genetically engineered glucose binding protein labelled with the environmentally sensitive dye badan. This system was developed in a search for an appropriate recognition-reporter unit to serve as a part of fluorescence-based sensor for continuous blood glucose monitoring. This labelled biomolecule represents an interesting subject for a single molecule study. Due to technical reasons however, single molecule spectroscopy could not be applied in this case. Therefore, conventional ensemble fluorescence spectroscopy methods were used to characterise behaviour of the labelled protein at different glucose concentrations. The last part of the thesis deals with instrumental aspects of single molecule imaging and spectroscopy. The aim of the work was to assess the applicability of a freshly installed commercial microscope a-SNOM (WITec GmbH) in single molecule fluorescence studies and at the same time to adopt the technique for future experiments in our research group.

This book is the first detailed and comprehensive guide to the theory of optical band shape of guest-molecule-doped crystals, polymers and glasses. Its main focus is on the dynamics of a single molecule, measured with the help of a train of photons emitted at random time moments.