Biophysical Society Thematic Meeting | Tutzing 2026

PROGRAM & ABSTRACTS

Biophysical Society Thematic Meetings

Single-Molecule FRET: The Next 30 Years

Tutzing, Germany | September 21–25, 2026

Thank you to the following supporters:

Thank you to the following sponsors:

Organizing Committee

Victoria Birkedal, Aarhus University, Denmark Don C. Lamb, Ludwig Maximilian University of Munich, Germany Claus Seidel, Heinrich Heine University Düsseldorf, Germany

Thank You to Our Supporters

Thank you to all supporters for their contributions.

Thank You to Our Sponsors

Thank you to all sponsors for their support.

Single-Molecule FRET: The Next 30 Years

Welcome Letter

September 2026

Dear Colleagues,

We would like to welcome you to the Biophysical Society Thematic Meeting entitled, Singe Molecule FRET: The Next 30 Years . In 1996, the first singe-molecule Förster Resonance Energy Transfer (smFRET) experiments were published, demonstrating that the possibility of measuring FRET efficiencies on individual molecules and opening a new direction for the biophysical sciences. With smFRET studies, heterogeneities can be observed directly and dynamics can be measured without synchronization. Since FRET is sensitive to distances, it is now possible to perform structural studies on dynamic systems and map the conformational landscape and functional heterogeneities of biomolecules under ambient conditions. In honor of this historic occasion, this conference will act as a forum to discuss the current advances and challenges for FRET experiments and theory. We hope to initiate visions for the future of smFRET studies and trigger further community activities. Overall, this conference features 73 posters, 54 lectures, and brings together over 145 scientists from a wide range of backgrounds and expertise. We hope that this meeting will not only provide a place to share your recent findings, but also to help promote new collaborations, helpful discussions, and future connections. We are also grateful to our generous sponsors, who have enabled us to honor exceptional contributions with a PicoQuant Early Career Award and Biophysical Journal Poster Awards. We invite you all to actively take part in the discussions following each talk, interact during the poster sessions, and exchanges thoughts and ideas in a relaxed environment during the coffee breaks and meals. We also hope that you will also enjoy the beautiful surroundings of Lake Starnberg and the town of Tutzing!

The Organizing Committee Victoria Birkedal

Don C. Lamb Claus Seidel

Single-Molecule FRET: The Next 30 Years

Meeting Code of Conduct

Biophysical Society Code of Conduct, Anti-Harassment Policy The Biophysical Society (BPS) is committed to providing an environment that encourages the free expression and exchange of scientific ideas. As a global, professional Society, the BPS is committed to the philosophy of equal opportunity and respectful treatment for all, regardless of national or ethnic origin, religion or religious belief, gender, gender identity or expression, race, color, age, marital status, sexual orientation, disabilities, veteran status, or any other reason not related to scientific merit. All BPS meetings and BPS-sponsored activities promote an environment that is free of inappropriate behavior and harassment by or toward all attendees and participants of Society events, including speakers, organizers, students, guests, media, exhibitors, staff, vendors, and other suppliers. BPS expects anyone associated with an official BPS-sponsored event to respect the rules and policies of the Society, the venue, the hotels, and the city. Definition of Harassment The term “harassment” includes but is not limited to epithets, unwelcome slurs, jokes, or verbal, graphic or physical conduct relating to an individual’s race, color, religious creed, sex, national origin, ancestry, citizenship status, age, gender or sexual orientation that denigrate or show hostility or aversion toward an individual or group. Sexual harassment refers to unwelcome sexual advances, requests for sexual favors, and other verbal or physical conduct of a sexual nature. Behavior and language that are welcome/acceptable to one person may be unwelcome/offensive to another. Consequently, individuals must use discretion to ensure that their words and actions communicate respect for others. This is especially important for those in positions of authority since individuals with lower rank or status may be reluctant to express their objections or discomfort regarding unwelcome behavior. It does not refer to occasional compliments of a socially acceptable nature. It refers to behavior that is not welcome, is personally offensive, debilitates morale, and therefore, interferes with work effectiveness. The following are examples of behavior that, when unwelcome, may constitute sexual harassment: sexual flirtations, advances, or propositions; verbal comments or physical actions of a sexual nature; sexually degrading words used to describe an individual; a display of sexually suggestive objects or pictures; sexually explicit jokes; unnecessary touching. Attendees or participants who are asked to stop engaging in harassing behavior are expected to comply immediately. Anyone who feels harassed is encouraged to immediately inform the alleged harasser that the behavior is unwelcome. In many instances, the person is unaware that their conduct is offensive and when so advised can easily and willingly correct the conduct so that it does not reoccur. Anyone who feels harassed is NOT REQUIRED to address the person believed guilty of inappropriate treatment. If the informal discussion with the alleged harasser is unsuccessful in remedying the problem or if the complainant does not feel comfortable with such an approach, they can report the behavior as detailed below. Reported or suspected occurrences of harassment will be promptly and thoroughly investigated. Following an investigation, BPS will immediately take any necessary and appropriate action. BPS will not permit or condone any acts of retaliation against anyone who files harassment complaints or cooperates in the investigation of same. Reporting a Violation Violations of this Conduct Policy should be reported immediately. If you feel physically unsafe or believe a crime has been committed, you should report it to the police immediately. To report a violation to BPS:

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Single-Molecule FRET: The Next 30 Years

Meeting Code of Conduct

 You may do so in person to BPS senior staff at Thematic Meetings, BPS Conferences, or other BPS events.

 At any time (during or after an event), you can make a report through

http://biophysics.ethicspoint.com or via a dedicated hotline (phone numbers listed on the website) which will collect and relay information in a secure and sensitive manner.

Reported or suspected occurrences of harassment will be promptly and thoroughly investigated per the procedure detailed below. Following an investigation, BPS will immediately take any necessary and appropriate action. BPS will not permit or condone any acts of retaliation against anyone who files harassment complaints or cooperates in the investigation of same. Investigative Procedure All reports of harassment or sexual harassment will be treated seriously. However, absolute confidentiality cannot be promised nor can it be assured. BPS will conduct an investigation of any complaint of harassment or sexual harassment, which may require limited disclosure of pertinent information to certain parties, including the alleged harasser. Once a complaint of harassment or sexual harassment is received, BPS will begin a prompt and thorough investigation. Please note, if a complaint is filed anonymously, BPS may be severely limited in our ability to follow-up on the allegation.  An impartial investigative committee, consisting of the current President, President-Elect, and Executive Officer will be established. If any of these individuals were to be named in an allegation, they would be excluded from the committee.  The committee will interview the complainant and review the written complaint. If no written complaint exists, one will be requested.  The committee will speak to the alleged offender and present the complaint.  The alleged offender will be given the opportunity to address the complaint, with sufficient time to respond to the evidence and bring his/her own evidence.  If the facts are in dispute, the investigative team may need to interview anyone named as witnesses.  The investigative committee may seek BPS Counsel’s advice.  Once the investigation is complete, the committee will report their findings and make recommendations to the Society Officers.  If the severity of the allegation is high, is a possible repeat offense, or is determined to be beyond BPS’s capacity to assess claims and views on either side, BPS may refer the case to the alleged offender’s home institution (Office of Research Integrity of similar), employer, licensing board, or law enforcement for their investigation and decision. Disciplinary Actions Individuals engaging in behavior prohibited by this policy as well as those making allegations of harassment in bad faith will be subject to disciplinary action. Such actions range from a written warning to ejection from the meeting or activity in question without refund of registration fees, being banned from participating in future Society meetings or Society-sponsored activities, being expelled from membership in the Society, and reporting the behavior to their employer or calling the authorities. In the event that the individual is dissatisfied with the results of the investigation, they may appeal to the President of the Society. Any questions regarding this policy should be directed to the BPS Executive Officer or other Society Officer.

Single-Molecule FRET: The Next 30 Years

Table of Contents

Table of Contents

General Information……………………………………………………………………………....1 Program Schedule..……………………………………………………………………………….4 Speaker Abstracts………………………………………………………………………………...11 Poster Sessions…………………………………………………………………………………...61

Single-Molecule FRET: The Next 30 Years

General Information

GENERAL INFORMATION

Registration/Information Location and Hours The meeting will take place at the Evangelische Akademie Tutzing, located at Schloßstraße 2+4, 82327 Tutzing, Germany. To pick up your badge and meeting materials, please visit the BPS Registration Desk, located in the Foyer during the following times: Monday, September 21 13:30 – 20:00 Tuesday, September 22 8:30 – 18:00 Wednesday, September 23 8:30 – 18:00 Thursday, September 24 8:30 – 16:00 Friday, September 25 8:30 – 13:00 Instructions for Presentations (1) Presentation Facilities: A data projector will be available in the Musiksall (Music Hall). Speakers are required to bring their own laptops and adaptors. It is recommended to have a backup of the presentation on a USB drive in case of any unforeseen circumstances. Speakers are advised to preview their final presentations before the start of each session. (2) Poster Session: 1) All poster sessions will be held in the Seminarraum, Medienraum, and Musiksaal (Seminar Room, Media Room, and Music Hall). 2) A display board measuring 1.2 meters wide by 1.5 meters high (portrait orientation) will be provided for each poster. Poster boards are numbered according to the same numbering scheme as listed in the E-book. 3) Typeface should be large enough to be read comfortably by interested attendees from distances of 1.5 meters (4-5 feet). Authors are expected to bring their own pushpins, thumbtacks, or Velcro for mounting poster materials. 4) There will be formal poster presentations on Tuesday and Thursday. Poster presentation times vary by day, so please refer to the program book for your formal presentation date and time. Two hours have been allotted for poster presentations each day. Presenting authors with odd-numbered poster boards should present during the first 60 minutes, and those with even-numbered poster boards should present during the last 60 minutes. Posters may only be displayed on the day of your assigned presentation and must be removed at the conclusion of your poster session. 5) During the assigned poster presentation sessions, presenters are requested to remain in front of their poster boards to meet with attendees. 6) All posters left uncollected at the end of the meeting will be discarded.

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Single-Molecule FRET: The Next 30 Years

General Information

Note Pads/Pens Society pens will be provided, however please bring your own note pad. Meals, Coffee Breaks, and Socials On the opening night, the welcome reception will be held in the Foyer and the Piano Concert will be held in the Musiksaal (Music Hall). All coffee breaks will be served in Foyer. From Tuesday–Thursday, lunches will be served in the Evangelische Akademie Restaurant. Wednesday evening dinner will also be held in the Evangelische Akademie Restaurant. Overflow seating will be available in the Kavaliersgewölbe (Cavalier Room). On Friday, a take-away boxed lunch will be provided in the Foyer from 11:00–13:00. The Thursday evening boat tour and banquet dinner will be held at 18:15 (6:15 PM) aboard the MS Starnberg. The dock is a short walk from the Akademie and directions will be provided onsite. The MS Starnberg is a non-smoking vessel. The Thursday evening dance will be held at 21:00 at the Akademie following the boat tour in the Schlossdiele (Castle Hall) & Salons. Housing at Evangelische Akademie Tutzing For attendees with sleeping accommodations at the Akademie, you may arrive at the Akademie after 11:00 AM on Monday, September 21. However, please be advised that a room may not be ready until the early afternoon. All guests must check out by 9:00 AM on Friday, September 25. Luggage storage is available for Akademie guests who require it upon check-in and/or check-out. Parking at Evangelische Akademie Tutzing Parking at the Akademie is limited and can only accommodate up to 40 vehicles daily. Additional local parking is available at the Tutzing Train Station (limited) and public parking areas for a fee. If you secured accommodations offsite, please consider alternative modes of transportation. Smoking Please be advised that the Evangelische Akademie Tutzing is a non-smoking facility. Name Badges Name badges will be given to you when you check in at the Registration Desk in the Foyer. Badges are required to enter all scientific sessions, poster sessions, and social functions. Please wear your badge throughout the conference. Internet Wi-Fi will be provided at the venue. Information will be available at the registration desk. On-Site Contact Information If you have any further requirements during the meeting, please contact the meeting staff at the registration desk from September 21–25 during registration hours.

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Single-Molecule FRET: The Next 30 Years

General Information

In case of emergency, you may contact the following: Dorothy Chaconas Phone: 301-785-0802 Email: dchaconas@biophysics.org Erica Bellavia Phone: 571-435-7669 Email: ebellavia@biophysics.org

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

Single-Molecule FRET: The Next 30 Years Tutzing, Germany September 21-25, 2026 All scientific sessions will be held in Musiksaal unless otherwise noted.

Monday, September 21, 2026

13:30 – 20:00

Registration & Information

Foyer

14:30 – 16:30

Introductory Workshops for FRET Tools 1. Deriving Theory for FRET 2. Understanding Analysis Tools

Seminarraum Musiksaal Kavaliersgewölbe

3. From the Design of FRET Assays to Structure Modelling

and Database Deposition

16:30 – 18:00

Welcome Reception

Foyer

Session I

Welcome Remarks & Keynote Talks Chair: Don C. Lamb, Ludwig Maximilian University of Munich, Germany Victoria Birkedal, Aarhus University, Denmark; Don C. Lamb, Ludwig Maximilian University of Munich, Germany Claus Seidel, Heinrich Heine University Düsseldorf, Germany Welcome & History of FRET Taekjip Ha, Harvard Medical School, USA Conformational Control: From Single-Molecule FRET to Novel Biotechnologies Philip Tinnefeld, Ludwig Maximilian University of Munich, Germany From Scaling Laws of Energy Transfer to FRET Sensing of Brownian DNA Computing

18:00 – 18:40

18:40 – 19:25

19:25 – 20:10

20:10 – 20:30

Networking Break

20:30 – 21:30

Piano Concert by Antonio Acunto

Musiksaal

Tuesday, September 22, 2026 8:30 – 18:00

Registration & Information

Foyer

Session II

Application of FRET in Biophysics Chair: Maria Ott, Martin Luther University Halle-Wittenberg, Germany Edward Lemke, Johannes Gutenberg University Mainz, Germany Decoding Molecular Plasticity in the Dark Proteome Jelle Hendrix, Hasselt University, Belgium Kinetic Modelling of Multi-State Protein Systems Using smFRET

9:00 – 9:30

9:30 – 10:00

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

10:00 – 10:20

Chara Sarafoglou, Institute of Molecular Biology and Biotechnology, Heraklion, Crete, Greece* , PQ-ECA smFRET-Guided Integrative Biophysics Reveals Allosteric Modules as Energetic Control Points of Protein Free-Energy Landscapes

10:20 – 10:50

Coffee Break

Foyer

Session III

Application of FRET in Proteins Chair: Gillian Cadden, Aarhus University, Denmark

10:50 – 11:20

Emmanuel Margeat, CNRS-Montpellier, France Looking at Metabotropic Glutamate Receptors Dynamics with Single-Molecule FRET Ecenaz Bilgen, Ludwig Maximilian University of Munich, Germany* , PQ-ECA Exploring Dynamic Protein Systems with Single-Molecule FRET Catherine Ghosh, University of California, Merced, USA* , PQ-ECA The Speed Limit of Genomic Search: A Kinetic Race Between Electrostatic Steering, Sliding Friction, and Conformational Locking Elizabeth Rhoades, University of Pennsylvania, Philadelphia, USA* Phosphoregulation of Tau Function

11:20 – 11:40

11:40 – 12:00

12:00 – 12:20

12:20 – 14:00

Lunch

Restaurant Evangelische Akademie

Session IV

Method Development / Probes Chair: Line Lund, Aarhus University, Denmark

14:00 – 14:30

Scott Blanchard, St. Judes Children’s Research Hospital, USA Quantitative Investigations of Complex Molecular Machines Using smFRET: What We Have Learned and Where Are We Going Allison H. Squires, University of Chicago, USA Design and Differentiation of FRET Constructs – Easy as ABEL-PIE Thomas-Otavio Peulen, Technical University Dortmund, Germany* From Structure to Observables: Optimal Dye Models for Predicting Lifetimes, Anisotropies, and FRET

14:30 – 15:00

15:00 – 15:20

15:20 – 15:50

Coffee Break

Foyer

Session V

Method Development Chair: Claus Seidel, Heinrich Heine University Düsseldorf, Germany

15:50 – 16:20

Rainer Erdmann, PicoQuant GmbH, Germany smFRET in Industry – A 35-Year Journey

16:20 – 16:50

Karl-Heinz Drexhage Memorial Lecture Jörg Enderlein, Georg August University, Göttingen, Germany The Electrodynamics of Fluorescing Molecules: A Tribute to Karl-Heinz Drexhage Eitan Lerner, The Hebrew University of Jerusalem, Israel* The “Bliss” of Observing Single-Molecule Bursts from Clusters in Live Cell Biomolecular Condensate

16:50 – 17:10

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

17:10 – 17:30

Quan Wang, NIH, Bethesda, MD, USA* Recent Developments of ABEL-FRET: Resolution Limits, Fast Dynamics and Biophysical Applications Mahran Shehade, Weizmann Institute of Science, Rehovot, Israel* , PQ-ECA Toward Understanding Structural Transitions and Dynamics in Proteins Using Plasmon-Enhanced smFRET Mirjam Kümmerlin, University of Oxford, United Kingdom* , PQ-ECA The Dark Side of FRET: Self-Quenching DNA Probes Enable Hour-Long smFRET and Live-Cell RNA Tracking

17:30 – 17:50

17:50 – 18:10

18:15 – 20:30

Poster Session

Seminarraum/Medienraum/Musiksaal

with Bavarian Brotzeit (Dinner) & Drinks

20:30 – 21:30

Parallel Round Table Discussions 1. FRET Standardization 2. FRET-Based Integrative Structural Biology 3. Women in FRET 4. Triggering Innovations in FRET

Room Assignments Made Onsite

Wednesday, September 23, 2026 8:30 – 18:00

Registration & Information

Foyer

Session VI

Application of FRET in Biochemistry Chair: Pooyeh Asadiatouei, Goethe University Frankfurt, Germany

9:00 – 9:45

Keynote Talk: Dagmar Klostermeier, University of Münster, Germany Deciphering the Mechanisms of Molecular Machines That Unwind and Untangle Nucleic Acids by Single-Molecule FRET: From Retrospective to Perspective Dina Grohman, University of Regensburg, Germany Single-Molecule Insights into the Dynamic Human RNA Interference Machinery Using SLAM-FRET Katherina Hemmen, University of Würzburg, Germany* Quantitative Image Spectroscopy for Protein-Protein Interactions in Living Cells: An Integrated FRET Framework

9:45 – 10:15

10:15 – 10:35

10:35 – 11:00

Coffee Break

Foyer

Session VI I

FRET in Cellular Experiments Chair: Jacob Piehler, Osnabrück University, Germany Thorsten Hugel, University of Freiburg, Germany* Single-Molecule FRET Trajectories in Living Cells

11:00 – 11:20

11:20 – 11:40

Sharonda LeBlanc, North Carolina State University, Raleigh, NC, USA* Live-Cell Flim-FRET Reveals Dynamic ATPase-Driven Pre-Ribosome Remodeling States

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

11:40 – 12:00

Nam Ki Lee, Seoul National University, South Korea* Direct Quantification of Protein-Protein Interactions in Living Bacterial Cells by Accurate FRET Measurement Greta Schmidt, Worcester Polytechnic Institute, MA, USA* , PQ-ECA Uncovering Novel G-Protein Pathways Using Live Cell Imaging Yunqing Li, Goethe University Frankfurt, Germany* , PQ-ECA Resolving MET Receptor Activation in Cells by Single-Molecule FRET

12:00 – 12:20

12:20 – 12:40

12:40 – 14:00

Lunch

Restaurant Evangelische Akademie

Session VIII

Theory and Analysis of FRET I Chair: Daniel Nettels, University of Zurich, Switzerland

14:00 – 14:30

Irina Gopich, NIDDK, NIH, Bethesda, MD, USA Photon-by-Photon Maximum Likelihood Analysis of Freely Diffusing Single Molecules Hagen Hofmann, Weizmann Institute of Science, Israel Model-Free Photon Analysis of Diffusion-Based Single-Molecule FRET Experiments Hoi Sung Chung, NIDDK, NIH, Bethesda, MD USA* Characterizing Ultrafast Barrier Crossing Dynamics of Protein Folding Using Two- and Three-Color Single-Molecule FRET in Zero-Mode Waveguides Lars Dingeldein, Frankfurt Institute of Advanced Studies, Goethe University Frankfurt, Germany* , PQ-ECA A Photon-by-Photon Likelihood for Continuous Free-Energy Landscapes and Diffusion Coefficients from Single-Molecule FRET Theory and Analysis of FRET II Chair: Tomoaki Yagi, RIKEN, Wako Campus, Saitama, Japan Gilad Haran, Weizmann Institute of Science, Rehovot, Israel Direct Observation of Translocation by a AAA + Machine Richard Börner, University of Applied Sciences Mittweida, Germany From FRET-Guided RNA 3D Structures to Conformational Ensembles Keynote Talks: smFRET, Back to the Future Chair: Don C. Lamb, Ludwig Maximilian University of Munich, Germany Don C. Lamb, Ludwig Maximilian University of Munich, Germany Summary of Round Table Discussions Ben Schuler, University of Zürich, Switzerland Probing the Rapid Interaction Dynamics of Charged Disordered Proteins Shimon Weiss, University of California, Los Angeles, USA spFRET/smFRET - A Story of Tools Making (and Their Utilization) Coffee & Networking Break Dinner

14:30 – 15:00

15:00 – 15:20

15:20 – 15:40

15:40 – 17:00

Foyer

Session IX

17:00 – 17:30

17:30 – 18:00

18:00 – 19:20

Restaurant Evangelische Akademie

Session X

19:20 – 19:40

19:40 – 20:25

20:25 – 21:10

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

21:10 – 21:40

Surprise

Musiksaal

Thursday, September 24, 2026 8:30 – 16:00

Registration & Information

Foyer

Session XI

Application of FRET in Structural Biology Chair: Thorben Cordes, Technical University Dortmund, Germany Sigrid Milles, Leibniz-FMP, Berlin, Germany Protein Disorder Across Scales – Combining NMR and Single-Molecule Fluorescence Nils-Alexander Lakomek, Forschungszentrum Jülich GmbH, Germany* Conformational Dynamics and Conformational Selection of Intrinsically Disordered SNAP25 During Snare Complex Assembly Hugo Sanabria, Clemson University, SC, USA* Frustrated Flexibility Shapes the Dynamics of Calmodulin Trapping Mikayel Aznauryan, Inserm, University of Bordeaux, France* Single-Molecule Spectroscopy of Disordered Protein Self-Assembly and RNA Recognition Viktorija Glembockyte, Max Planck Institute for Medical Research, Heidelberg, Germany Programming FRET: Engineering Precise Interactions for Modular Sensing and Superresolution Imaging Andrea Soranno, Washington University in St. Louis, USA* Single-Molecule Spectroscopy of Disordered Regions and Nucleic Acids in Bacterial Transcription Jagadish Hazra, Indian Institute of Science Education and Research, Mohali, India* , PQ-ECA Elucidating Single-Molecule Reaction Kinetics Using Multiplexed in Situ Fluorescence-Based DNA Sequencing Hannah Baird, Cardiff University, United Kingdom* , PQ-ECA Single-Molecule FRET Insights into the Dynamic Synergistic Mechanism of Antimicrobial Peptides Coffee Break FRET: From Methods to Applications Chair: Christian Hübner, University of Lübeck, Germany

9:00 – 9:30

9:30 – 9:50

9:50 – 10:10

10:10 – 10:30

10:30 – 10:50

Foyer

Session XII

10:50 – 11:20

11:20 – 11:40

11:40 – 12:00

12:00 – 12:20

12:20 – 14:00

Lunch

Restaurant Evangelische Akademie

Session XIII

FRET: Studies of Protein Nucleic Acid Interactions Chair: Michael Schlierf, Dresden University of Technology, Germany

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

14:00 – 14:20

Nils Walter, University of Michigan, Ann Arbor, MI. USA* Flux and Function: Watching RNA-Protein Complexes Assemble to Control Gene Expression Nusrat Qureshi, European Molecular Biology Laboratory, Heidelberg, Germany* , PQ-ECA Sync or Sink: The Extent of Transcription-Translation Coupling Determines Transcription Factor Recruitment to Regulate Gene Expression Johannes Hohlbein, Brandenburg University of Technology Cottbus-Senftenberg, Germany* DNA FRET Sensors: From Probing DNA Synthesis in Nanofluidic Devices to Quantifying Complex Interactions Between DNA Transcription Factors Andreas Hartmann, Dresden University of Technology, Germany* Different Personalities of Phase-Separating Proteins Reveal Multiple RNA Binding Modes

14:20 – 14:40

14:40 – 15:00

15:00 – 15:20

Mariska Haas, Ulm University, Germany* , PQ-ECA Quantitative smFRET of Low-Affinity Complexes in Nanoholes

15:20 – 15:40

15:40 – 16:00

Yale Goldmann, University of California, Davis, USA* Nanometer-Scale RNA Protein Clusters (RPCS) Foster Helicase Activity of Dead Box Helicase EIF4A

16:00 – 18:00

Poster Session & Coffee Break

Seminarraum/Medienraum/Musiksaal

18:15 – 21:00

Boat Tour & Banquet Dinner

MS Starnberg on Lake Starnberg

21:00

Music and Dance: TUM Old's Cool Combo

Schlossdiele & Salons

Friday, September 25, 2026 8:30 – 13:00

Registration & Information

Foyer

Session XIV

New Directions in FRET I Chair: Georg Krainer, University of Graz, Austria

9:00 – 9:30

Sebastian Deindl, Uppsala Biomedical Center, Sweden From Sequence to Function: Bridging Single-Molecule Kinetics with Molecular Identity Sonja Schmid, University of Basel, Switzerland Breaking the Photobleaching Limit in Single-Molecule FRET by Dyecycling Tim Schröder, Ludwig Maximilian University of Munich, Germany* Microtime-Gated Photon Statistics: From Fast FRET Dynamics to Exciton Colliders

9:30 – 10:00

10:00 – 10:20

10:20 – 10:50

Coffee Break

Foyer

Session XV

New Directions in FRET II Chair: Victoria Birkedal, Aarhus University, Denmark

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Single-Molecule FRET: The Next 30 Years

Daily Schedule

10:50 – 11:10

Evelyn Ploetz, Ludwig Maximilian University of Munich, Germany* Combining Single-Molecule FRET and Graphene Energy Transfer to Resolve Promoter-Dependent DNA Deformation by TBP Chirlmin Joo, Delft University of Technology, The Netherlands Next-Generation Single-Molecule Tools for Exploring Biomolecular Sequence Space Keynote Talk: Claus Seidel, Heinrich Heine University Düsseldorf, Germany Dynamic Structural Biology with smFRET and FRET Nanoscopy Victoria Birkedal, Aarhus University, Denmark Don C. Lamb, Ludwig Maximilian University of Munich, Germany Claus Seidel, Heinrich Heine University Düsseldorf, Germany Closing Remarks & Presentation of PicoQuant Early Career Award and BJ Poster Awards

11:10 – 11:40

11:40 – 12:25

12:25 – 12:45

11:00 – 13:00

Boxed Lunches

Foyer

* Contributed talks selected from among submitted abstracts PQ-ECA Contributed talks eligible for the PicoQuant Early Career Award

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Single-Molecule FRET: The Next 30 Years

Speaker Abstracts

SPEAKER ABSTRACTS

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Single-Molecule FRET: The Next 30 Years

Monday Speaker Abstracts

CONFORMATIONAL CONTROL: FROM SINGLE MOLECULE FRET TO NOVEL BIOTECHNOLOGIES Taekjip Ha ;

1 Boston Children's Hospital, Boston, MA, USA 2 Harvard Medical School, Boston, MA, USA

After an introduction to single-molecule FRET from my own personal perspective, I will discuss how it can be used to uncover functional regulation via conformational control and how we leverage these fundamental insights to engineer powerful biotechnologies. For example, our single-molecule investigations into CRISPR-Cas9 DNA unwinding and conformational dynamics have yielded a suite of tools. By exploiting the differential sequence requirements for Cas9 binding versus cleavage, we engineered a light-controlled, very fast CRISPR (vfCRISPR) system to synchronously generate double-strand breaks (DSBs) across cell populations. Tracking these repair kinetics in real time has provided fundamental insights into nonhomologous end joining and homologous recombination. Concurrently, we developed GOLDFISH for high resolution, non-denaturing genome imaging and successfully integrated it with vfCRISPR. Beyond CRISPR, I will introduce SHARP, a novel isothermal DNA amplification technology for repetitive sequences, which emerged directly from our foundational studies on helicase regulation and the subsequent engineering of superhelicases. Finally, I will present single molecule FRET investigations of prime-editing mechanisms, in which we dissect the individual steps of this gene-editing method without generating a DSB. FROM SCALING LAWS OF ENERGY TRANSFER TO FRET SENSING OF BROWNIAN DNA COMPUTING Philip Tinnefeld 1 ; 1 Ludwig-Maximilians-Universität München, Chemistry, Munich, Germany Förster resonance energy transfer obeys the famous r -6 dependence for the interaction of two dipoles in the near-field. Using DNA nanotechnology, we arranged dye molecules relative to different materials such as gold nanoparticles, carbon nanotubes and graphene and studied the scaling laws experimentally. Among the different options, graphene energy transfer (GET) in combination with the discovery of a vertical orientation of dsDNA on graphene turned out to be most useful and has emerged into a versatile technique for studying DNA-protein interactions with nanometer precise superresolution microscopy. Here, the axial information is deduced from GET-efficiency and xy-information is obtained by photon-efficient pMINFLUX revealing simultaneous bending and torsion of enzyme induced DNA conformations. Finally, we discuss how mutli-color smFRET with dark quenchers probes states of DNA nanorobots and Brownian DNA computers that work close to the thermodynamic optimum.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

DECODING MOLECULAR PLASTICITY IN THE DARK PROTEOME Edward A. Lemke 1,2 ; 1 JGU, Biology & Chemistry, Mainz, Germany 2 IMB, IMB, Mainz, Germany

The mechanisms by which intrinsically disordered proteins (IDPs) engage in rapid and highly selective binding is a subject of considerable interest and represents a central paradigm to nuclear pore complex (NPC) function. Nuclear transport receptors (NTRs) can move through the central channel of the NPC which is filled with hundreds of phenylalanine-glycine-rich nucleoporins (FG-Nups) reaching millimolar concentrations with elusive conformational plasticity. Since site specific labeling of proteins with small but highly photostable fluorescent dyes inside cells remains the major bottleneck for directly studying protein dynamics in the cellular interior, we have now developed a semi-synthetic strategy based on novel artificial amino acids that are easily and site-specifically introduced into any protein by the natural machinery of the living cell via a newly developed thin-film synthetic organelle that equips the living cell with up to three genetic codes. This allowed us to develop an experimental approach combining site-specific fluorescent labeling of IDPs in non-fixed cells with fluorescent lifetime imaging microscopy (FLIM) to directly decipher the plasticity of FG-Nups via FRET. Our study enabled a conformational look on the condensated IDPs in the sub-resolution (roughly (50 nm) 3 small cavity) cavity of the NPC. By measuring the end-to-end distances of different segments of the labeled FG-Nups using time resolved scanning FRET and anisotropy spectroscopy, we can extract the scaling exponent and dynamics, which directly describes the conformations of FG Nups at their functional status as well as the solvent quality in the cellular and even inner NPC environment.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

METHODOLOGIES FOR FLUORESCENCE BASED STRUCTURAL BIOLOGY OF COMPLEX SYSTEMS Jelle Hendrix 1 ; 1 Hasselt University, Hasselt, Belgium Biomolecular function is inherently dynamic, and resolving how structural fluctuations shape activity requires experimental approaches capable of capturing motion across molecular, spatial, and temporal scales. We employ single-molecule FRET (smFRET) to detect rapid conformational changes within individual molecules, even when state switching occurs during millisecond bursts, providing direct access to kinetic information. A major focus is the development of physiologically relevant model environments. Using GUVs, supported bilayers, and pore-spanning membranes, combined with two-photon polymerization, we fabricate custom 3D microstructures compatible with confocal and multiphoton microscopes. Microfluidic devices enable controlled flow, droplet manipulation, and long-duration smFRET, supported by array-detector FCS and 3D flow-velocity mapping. Finally, resistive microheating and lock-in detection allow precise thermal perturbations for extracting kinetic parameters, while solvatochromic dyes (e.g., ACDAN) report on local environments through spectral and lifetime phasors. Together, these methods form a versatile, microscope-integrated toolkit for quantifying biomolecular dynamics in tunable microenvironments.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

SMFRET-GUIDED INTEGRATIVE BIOPHYSICS REVEALS ALLOSTERIC MODULES AS ENERGETIC CONTROL POINTS OF PROTEIN FREE-ENERGY LANDSCAPES Chara Sarafoglou 1,2 ; Yusran A Muthahari 1 ; Charalambos Pozidis 1 ; Andreas Kofidis 3 ; Mary Providaki 1 ; Alexis Molfetas 1 ; Mikis Mylonakis 4 ; Kostas Mavrakis 4 ; Reza Aditama 5 ; Rukman Hertadi 5 ; Giannis Zacharakis 4 ; Yannis Pantazis 3 ; Giorgos Gouridis 1 ; 1 Institute of Molecular Biology and Biotechnology , Structural Biology and Biophysics, Heraklion, Greece 2 University of Crete, Biology, Heraklion, Greece 3 Institute of Applied and Computational Mathematics, Heraklion, Greece 4 Institute of Electronic Structure and Lasers, Biophotonics and molecular imaging, Heraklion, Greece 5 Bandung Institute of Technology, Biochemistry, Bandung, Indonesia Free-Energy Landscapes (FELs) provide a unifying physical framework for understanding protein folding, allostery, ligand recognition, and functional trade-offs that underlie evolution. Yet a challenge remains: which structural and energetic units sculpt these landscapes, and how does their perturbation redirect protein behaviour across equilibrium and nonequilibrium biochemical processes? In my PhD, I addressed this challenge using substrate-binding proteins (SBPs) providing tractable model landscapes and single-molecule Förster Resonance Energy Transfer (smFRET) as the central experimental readout of conformational-state populations, transitions, and fluxes. Across folding, ligand binding, transport-coupled function, and evolutionary adaptation, a common principle emerged: coevolving allosteric modules act as energetic control points that define the wells, barriers, and connectivity of the FEL.By combining smFRET with time-varying mixture modeling in the NEXT-FRET framework (PNAS, doi:10.1073/pnas.2529979123), we tracked nonequilibrium population flow during folding of freely diffusing SBPs and resolved transient on-pathway intermediates. Rather than representing exclusively folding species, these intermediates correspond to wells within the same modular FEL that governs native-state dynamics and allostery. Signal peptides and chaperones reroute folding by selectively stabilizing these distinct intermediates and reshaping kinetic barriers. The same modules then fine-tune the native landscape to control function and evolvability. Integration with hydrogen–deuterium exchange mass spectrometry, molecular dynamics simulations, calorimetry, and transport-coupled assays revealed that perturbing them reweights conformational ensembles, switches binding mechanisms among induced fit, lock-and-key, and conformational selection, and tunes thermodynamically imposed functional trade-offs (bioRxiv, doi:10.1101/2025.10.08.680613). Extending this logic across related SBPs showed that affinity and specificity cannot be explained by binding-cleft chemistry alone. Instead, they emerge from coupling between local ligand contacts and the conformational free energy encoded by the allosteric network, providing a route to engineer protein function by reweighting pre-existing states rather than redesigning binding sites.Together, these studies unify folding, dynamics, function, and adaptation within a shared FEL framework.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

LOOKING AT METABOTROPIC GLUTAMATE RECEPTORS DYNAMICS WITH SINGLE MOLECULE FRET Emmanuel Margeat 1 ; 1 Centre de Biologie Structurale, Univ. Montpellier, CNRS, INSERM, Montpellier, France Metabotropic glutamate receptors (mGluR) regulate neuronal excitability and synaptic transmission by sensing L-glutamate – the major excitatory neurotransmitter in the central nervous system. Their crucial role for synaptic function makes them attractive targets for the treatment of numerous neurological and psychiatric diseases including for instance anxiety, depression, schizophrenia and addiction. To explore mGluR activation, we use single molecule FRET, that allows to screen the conformations explored by single protein complexes, with high temporal and spatial resolution. 3 color single-molecule FRET extends the conformational analysis to the measurements of 3 distances simultaneously, and therefore to the observation of correlated movements. However, the site-specific labeling of a biomolecular complex with 3 single molecule-compatible fluorophores remains challenging. We established a series of 2-color and 3-color smFRET sensors, through incorporation of up to two orthogonally reactive non-canonical amino-acids (ncAA) , and/or the addition of a SNAP self-labeling tag. These sensors report on the initial steps of mGluR2 activation, including the reorientation of the upper and the lower lobes of the Venus flytrap domain (VFT) or the cysteine rich domain (CRD) in an intersubunit fashion, the VFT closure in an intrasubunit fashion, and the correlation between these movements. We then used 2- and 3-color single molecule FRET to explore ligand induced conformational changes on mGlu2 homodimers. We show that agonist-binding efficiently depopulates the inactive state, leading to an equilibrium of receptors switching between the active and a newly identified intermediate state. Only the addition of a synthetic allosteric modulator, or of the G-protein, leads to a full stabilization of the activated receptor. We also extend this approach to the study of mglu2-containg heterodimers, offering new insights on the allosteric control of one protomer by the other, and on the asymmetric interaction with the G-proteins.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

EXPLORING DYNAMIC PROTEIN SYSTEMS WITH SINGLE-MOLECULE FRET Ecenaz Bilgen 1 ; Carina Fernandez Gonzalez 2 ; Léo Bonhomme 4 ; Mariam Mohamadi 3 ; Franz Hagn 3 ; Emmanuel Margeat 4 ; Robert B. Quast 4 ; Johannes Buchner 2 ; Don C. Lamb 1 ; 1 LMU Munich, Department of Chemistry and Center for Nanoscience, Munich, Germany 2 Technical University of Munich, Center for Functional Protein Assemblies (CPA), Department Bioscience, School of Natural Sciences, Munich, Germany 3 Technical University of Munich, Bavarian NMR Center (BNMRZ) and Structural Membrane Biochemistry, Dept. of Bioscience, TUM School of Natural Sciences, Munich, Germany 4 University of Montpellier, Centre de Biologie Structurale (CBS), CNRS, INSERM , Montpellier, France Single-molecule fluorescence resonance energy transfer (smFRET) has become a powerful approach for resolving the structure, dynamics, and interactions of biomolecules. This work presents a comprehensive investigation of dynamic protein systems using two- and three-color smFRET, with a particular focus on the solution-based multiparameter fluorescence detection with pulsed interleaved excitation (MFD-PIE) method. By combining fluorescence lifetime analysis, photon distribution analysis (PDA), and filtered fluorescence correlation spectroscopy (fFCS), conformational dynamics on the sub-millisecond timescale can be quantitatively characterized. The first part of this work investigates the conformational regulation of the tumor suppressor protein p53 by small heat shock proteins (sHsps). We show that α A- and α B crystallin (HspB4/HspB5) selectively bind p53 and trap it in a heterogeneous ensemble of non native conformations rather than a single defined state. We used FCS, PDA, and fluorescence lifetime analysis to elucidate the conformational changes occurring in the DNA-binding domain of p53 as it is released from the α B-crystallin complex. We demonstrate that p53 can be transferred to the ATP-dependent Hsp70/Hsp90 chaperone machinery for release and reactivation. The second part of this work demonstrates the application of two- and three-color smFRET to G protein-coupled receptor (GPCR) systems. By combining two-color smFRET with filtered fluorescence correlation spectroscopy (fFCS), we characterize the conformational changes and sub-millisecond dynamics of a transmembrane receptor. We further employ three color smFRET to investigate the human metabotropic glutamate receptor 2 (mGlu2), resolving coordinated conformational motions across multiple domains of the receptor. Together, these studies highlight the versatility of advanced smFRET methodologies for probing the complex conformational landscapes and dynamics of biologically important membrane proteins.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

THE SPEED LIMIT OF GENOMIC SEARCH: A KINETIC RACE BETWEEN ELECTROSTATIC STEERING, SLIDING FRICTION, AND CONFORMATIONAL LOCKING Catherine Ghosh 1,2 ; Rama R Goluguri 3 ; Mourad Sadqi 2 ; Victor Muñoz 2 ; 1 University of Copenhagen, Department of Biology, Copenhagen, Denmark 2 University of California Merced, Department of Bioengineering, Merced, CA, USA 3 Stanford University, Department of Biochemistry, Palo Alto, CA, USA Transcription factors (TFs) navigate the vast eukaryotic genome to locate specific target sites with remarkable speed and precision. While facilitated diffusion via 1D sliding is a widely accepted search mechanism, the precise physical principles governing the transition from rapid scanning to stable recognition remain unresolved. Specifically, the interplay between long-range electrostatic steering, the "friction" of sliding, and the conformational changes required for specific binding is poorly understood. In this work, we resolve this search mechanism for the Engrailed homeodomain (enHD) using advanced single-molecule FRET (smFRET) spectroscopy and Maximum Likelihood Analysis of Photon Arrival Times (MLA-PAT). By dissecting the binding kinetics across a broad range of ionic strengths and DNA lengths, we successfully extricated the distinct contributions of bulk electrostatics, local DNA field effects, and 1D sliding dynamics. Our results reveal that the search process is a finely tuned kinetic race. We experimentally determined the Debye length of the DNA electric field (0.82 nm at physiological ionic strength), confirming that electrostatic steering accelerates the initial capture. However, we uncovered a critical "sliding bottleneck": while longer DNA antennas enhance protein recruitment, the time cost of sliding eventually hampers the specific "lock-into-target" (cognate site binding) step. Mechanistically, we show that specific recognition is not merely diffusion arrest but is gated by a rate-limiting conformational switch. Crucially, we find that the DNA’s local electric field acts as an active catalyst by accelerating the protein's conformation switch. This switch acts as a "molecular brake," rheostatically tuned by the protein’s stability to halt diffusion precisely at the cognate site. This establishes a quantitative framework for understanding how eukaryotic TFs optimize the trade-off between search speed and recognition fidelity, avoiding the "stickiness" of non-specific traps while ensuring reliable gene activation.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

PHOSPHOREGULATION OF TAU FUNCTION Elizabeth Rhoades 1,2 ; Christopher R Brue 1,2 ; Yujing Fan 1 ; Sarah A Schick 1 ; Polina Holubovska 1 ; 1 University of Pennsylvania, Chemistry, Philadelphia, PA, USA 2 University of Pennsylvania Perelman School of Medicine, Biochemistry & Biophysics, Philadelphia, PA, USA The intrinsically disordered protein Tau plays an important role in modulating the dynamics and stability of axonal microtubules in neurons. Tubulin binding sites are found in Tau's proline-rich region (PRR), microtubule binding repeats (MTBRs), and pseudo-repeat (R'). Tau phosphorylation sites, which cluster with high frequency within the PRR, differentially regulate tubulin interactions although the molecular details are poorly. Here, we use single molecule Förster resonance energy transfer to probe the impact of phosphorylation on long range intramolecular interactions within Tau. We correlate these with tubulin binding and polymerization capacity to provide mechanistic insight into regulation of Tau function.

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Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

QUANTITATIVE INVESTIGATIONS OF COMPLEX MOLECULAR MACHINES USING SMFRET: WHAT WE HAVE LEARNED AND WHERE ARE WE GOING Scott C. Blanchard ; 1 St. Jude Children's Research Hospital, Structural Biology, Chemical Biology & Therapeutics, Memphis, TN, USA Static structures determined by crystallography and cryo-EM have transformed our understanding of the molecules of life. The power of single-molecule methods is to add the dimension of dynamism to these snapshots to reveal the nature, order and timing of structural processes underpinning function. For over two decades our group has worked alongside structural biologists, cell biologists, and pharmacologists to examine biomolecular metastability and how transitions between states are impacted by disease mutations and therapeutic interventions. Single-molecule FRET has been central to this pursuit. By resolving conformational dynamics one molecule at a time, it becomes possible to identify rate-limiting steps in function across machines spanning many orders of magnitude in mass, and to see how mutations and small molecules reshape energy landscapes in ways that static views cannot reveal. This mechanistic layer is where much of the molecular basis of human disease ultimately resides. Alongside advances across the field, our team has focused on widening the resolution, sensitivity and reproducibility of single-molecule fluorescence measurements. Hidden Markov approaches, adapted from the ion-channel community, brought statistical rigor to the interpretation of conformational trajectories; sCMOS detection improved throughput and time resolution; self-healing fluorophores extended the sensitivity, accuracy, and observation times attainable; and parallelized, rapid-exchange measurement suppresses experimental variances to reveal functional distinctions between closely related systems. Together with findings from many laboratories, our results make an increasingly clear case that dynamism is fundamental to biomolecular function and governing dynamism is essential to regulation. Understanding the evolutionary constraints that shape and tune the dynamic properties of biomolecules is a critical and largely unexplored frontier on the road ahead. Efforts toward this frontier will be discussed alongside new findings that naturally occurring sequence variation, chemical modification, and cellular context tune the dynamic landscapes of biomolecular machines and how these features may be understood and therapeutically engaged.

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