Related Experiment Video
Updated: May 24, 2026
08:21
Synthesis of In37P20(O2CR)51 Clusters and Their Conversion to InP Quantum Dots
Published on: May 7, 2019
Single-cluster pull-down assay for visualization and characterization of nanoclusters and condensates
1Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.
STAR Protocols
|May 22, 2026
Summary
Researchers developed a new method, single-cluster pull-down (SiCluP), to study how biomolecular condensates form. This fluorescence microscopy technique allows detailed analysis of early assembly stages and molecular dynamics within individual nanoclusters.
Area of Science:
- Cell Biology
- Biophysics
- Biochemistry
Background:
- Biomolecular condensates are crucial for cellular organization.
- Understanding the early stages of condensate assembly is challenging.
- Existing methods struggle to resolve dynamic processes at the single-nanocluster level.
Purpose of the Study:
- To introduce a novel fluorescence microscopy assay, single-cluster pull-down (SiCluP).
- To enable quantitative analysis of early biomolecular condensate assembly intermediates.
- To provide a method for studying nanocluster dynamics under controlled conditions.
Main Methods:
- Development of the SiCluP assay for selective nanocluster immobilization.
- Utilizing antibody-functionalized PEG surfaces for attachment.
- Employing Total Internal Reflection Fluorescence (TIRF) microscopy for imaging.
- Detailed protocol for surface preparation, nanocluster assembly, and time-resolved measurements.
Main Results:
- SiCluP allows selective immobilization of nanoclusters.
- The assay enables quantitative analysis of single nanoclusters.
- Demonstrated quantification of assembly kinetics, relative cluster size, and molecular exchange using FUS as a model system.
Conclusions:
- SiCluP is an effective method for studying early biomolecular condensate assembly.
- The technique provides unprecedented resolution for single-cluster analysis.
- This assay advances the understanding of dynamic processes in cellular organization.
