Krishnarao Appasani is the Founder and Chief Executive Officer of GeneExpression Systems, a global conference-producing organisation focusing on biomedical and physical sciences. He is also an elected Life Fellow of the Royal Society of Biologists, London, UK. Raghu Kiran Appasani is a Psychiatrist and Neuroscientist. He practices at University of Southern California. He is also the Founder and Chief Executive Officer of The MINDS Foundation.

'Single-molecule experiments have revolutionized our approaches to biophysics and biochemistry at molecular scales. Only a handful of reference textbooks serves the ever-expanding community of practitioners, beginners, and curious amateurs of single-molecule techniques. This volume is poised to become a new single-molecule reference work, to be readily at hand in any bioscience laboratory, to be read and understood by any scientist practicing, or simply interested, in the new world of experiments, reasoning, and statistical analysis opened up by single-molecule science.' Michel A. G. J. Orrit, Professor of Spectroscopy of Molecules in Condensed Matter, Leiden University, The Netherlands

• Preface; I. Super Resolution Microscopy and Molecular Imaging Techniques to Probe Biology: 1. Introduction on Single Molecule Science Krishnarao Appasani and Raghu K. Appasani; 2. One molecule, two molecules, red molecules, blue molecules: Methods for quantifying localization microscopy data George Patterson and Gaetan G. Herbomel; 3. Muti-scale fluorescence imaging Ruben Bulkescher, Inn Chung, Jan Philipp Eberle, Holger Erfle, Manuel Gunkel, Jürgen Reymann, Karsten Rippe, Guido Sauter, Ronald Simon and Vytaute Starkuviene; 4. Long-read single-molecule optical maps Assaf Grunwald, Yael Michaeli and Yuval Ebenstein; II. Protein Folding, Structure, Confirmation and Dynamics: 5. Single-molecule mechanics of protein nanomachines Gabriel Žoldák and Katarzyna Tych; 6. Post-translational Protein Translocation Through Membranes at the Single Molecule Level Diego Quiroga-Roger, Hilda M. Alfaro-Valdés and Christian A.M. Wilson Moya; III. Mapping DNA Molecules at Single Molecule Level: 7. Observing Dynamic states of single molecule DNA and Proteins using Atomic Force Microscope Jingqiang Li, Sithara Wijeratne and Ching-Hwa Kiang; 8. Atomic Force Microscopy and Detecting a DNA Biomarker of a Few Copies without Amplification Sourav Mishra, Yoonhee Lee and Joon Won Park; IV. Single Molecule Biology to Study Gene Expression: 9. Single molecule detection in the study of gene expression Vipin Kumar, Simon Leclerc and Yuichi Taniguchi.

Krishnarao Appasani is the Founder and Chief Executive Officer of GeneExpression Systems, a global conference-producing organisation focusing on biomedical and physical sciences. He is also an elected Life Fellow of the Royal Society of Biologists, London, UK. Raghu Kiran Appasani is a Psychiatrist and Neuroscientist. He practices at University of Southern California. He is also the Founder and Chief Executive Officer of The MINDS Foundation.

'Single-molecule experiments have revolutionized our approaches to biophysics and biochemistry at molecular scales. Only a handful of reference textbooks serves the ever-expanding community of practitioners, beginners, and curious amateurs of single-molecule techniques. This volume is poised to become a new single-molecule reference work, to be readily at hand in any bioscience laboratory, to be read and understood by any scientist practicing, or simply interested, in the new world of experiments, reasoning, and statistical analysis opened up by single-molecule science.' Michel A. G. J. Orrit, Professor of Spectroscopy of Molecules in Condensed Matter, Leiden University, The Netherlands

• Preface; I. Super Resolution Microscopy and Molecular Imaging Techniques to Probe Biology: 1. Introduction on Single Molecule Science Krishnarao Appasani and Raghu K. Appasani; 2. One molecule, two molecules, red molecules, blue molecules: Methods for quantifying localization microscopy data George Patterson and Gaetan G. Herbomel; 3. Muti-scale fluorescence imaging Ruben Bulkescher, Inn Chung, Jan Philipp Eberle, Holger Erfle, Manuel Gunkel, Jürgen Reymann, Karsten Rippe, Guido Sauter, Ronald Simon and Vytaute Starkuviene; 4. Long-read single-molecule optical maps Assaf Grunwald, Yael Michaeli and Yuval Ebenstein; II. Protein Folding, Structure, Confirmation and Dynamics: 5. Single-molecule mechanics of protein nanomachines Gabriel Žoldák and Katarzyna Tych; 6. Post-translational Protein Translocation Through Membranes at the Single Molecule Level Diego Quiroga-Roger, Hilda M. Alfaro-Valdés and Christian A.M. Wilson Moya; III. Mapping DNA Molecules at Single Molecule Level: 7. Observing Dynamic states of single molecule DNA and Proteins using Atomic Force Microscope Jingqiang Li, Sithara Wijeratne and Ching-Hwa Kiang; 8. Atomic Force Microscopy and Detecting a DNA Biomarker of a Few Copies without Amplification Sourav Mishra, Yoonhee Lee and Joon Won Park; IV. Single Molecule Biology to Study Gene Expression: 9. Single molecule detection in the study of gene expression Vipin Kumar, Simon Leclerc and Yuichi Taniguchi.

Krishnarao Appasani is the Founder and Chief Executive Officer of GeneExpression Systems, a global conference-producing organisation focusing on biomedical and physical sciences. He is also an elected Life Fellow of the Royal Society of Biologists, London, UK. Raghu Kiran Appasani is a Psychiatrist and Neuroscientist. He practices at University of Southern California. He is also the Founder and Chief Executive Officer of The MINDS Foundation.

'Single-molecule experiments have revolutionized our approaches to biophysics and biochemistry at molecular scales. Only a handful of reference textbooks serves the ever-expanding community of practitioners, beginners, and curious amateurs of single-molecule techniques. This volume is poised to become a new single-molecule reference work, to be readily at hand in any bioscience laboratory, to be read and understood by any scientist practicing, or simply interested, in the new world of experiments, reasoning, and statistical analysis opened up by single-molecule science.' Michel A. G. J. Orrit, Professor of Spectroscopy of Molecules in Condensed Matter, Leiden University, The Netherlands

• Preface; I. Super Resolution Microscopy and Molecular Imaging Techniques to Probe Biology: 1. Introduction on Single Molecule Science Krishnarao Appasani and Raghu K. Appasani; 2. One molecule, two molecules, red molecules, blue molecules: Methods for quantifying localization microscopy data George Patterson and Gaetan G. Herbomel; 3. Muti-scale fluorescence imaging Ruben Bulkescher, Inn Chung, Jan Philipp Eberle, Holger Erfle, Manuel Gunkel, Jürgen Reymann, Karsten Rippe, Guido Sauter, Ronald Simon and Vytaute Starkuviene; 4. Long-read single-molecule optical maps Assaf Grunwald, Yael Michaeli and Yuval Ebenstein; II. Protein Folding, Structure, Confirmation and Dynamics: 5. Single-molecule mechanics of protein nanomachines Gabriel Žoldák and Katarzyna Tych; 6. Post-translational Protein Translocation Through Membranes at the Single Molecule Level Diego Quiroga-Roger, Hilda M. Alfaro-Valdés and Christian A.M. Wilson Moya; III. Mapping DNA Molecules at Single Molecule Level: 7. Observing Dynamic states of single molecule DNA and Proteins using Atomic Force Microscope Jingqiang Li, Sithara Wijeratne and Ching-Hwa Kiang; 8. Atomic Force Microscopy and Detecting a DNA Biomarker of a Few Copies without Amplification Sourav Mishra, Yoonhee Lee and Joon Won Park; IV. Single Molecule Biology to Study Gene Expression: 9. Single molecule detection in the study of gene expression Vipin Kumar, Simon Leclerc and Yuichi Taniguchi.