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Best practice mass photometry: a guide to optimal single-molecule mass measurement

Abstract:
Mass photometry (MP) has emerged as a powerful approach to study quaternary biomolecular structure, dynamics and interactions. The capabilities of the method ultimately hinge on the ability to accurately measure the tiny optical contrast generated by individual molecules at a glass–water interface, which enables mass-resolved quantification of biomolecular mixtures. Ideally, this capability is limited only by photon shot noise, but in practice depends on additional parameters and details of the assay. Here, we focus on the key factors affecting MP performance and present simple steps that can be taken to achieve optimal MP measurements in terms of mass resolution, quantitative detection limit, reproducibility and analyte concentration range without compromising the speed and simplicity of the technique. Each sample takes <10 min to analyse, with an additionial 2 h if amination of the glass surface is desired.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/s41596-025-01255-4

Authors

More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Physiology Anatomy and Genetics
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Computer Science
Sub department:
Physics - Central
Oxford college:
St John's College
Role:
Author
ORCID:
0000-0001-8941-3693
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Chemistry
Role:
Author
ORCID:
0000-0002-6638-6298
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Physiology Anatomy and Genetics
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Physiology Anatomy and Genetics
Role:
Author


More from this funder
Funder identifier:
https://ror.org/029chgv08
Grant:
218514/Z/19/Z
More from this funder
Funder identifier:
https://ror.org/0472cxd90
Grant:
819593
More from this funder
Funder identifier:
https://ror.org/001aqnf71
Grant:
EP/X025713/1
More from this funder
Funder identifier:
https://ror.org/0439y7842
Grant:
EP/T03419X/1
More from this funder
Funder identifier:
https://ror.org/00cwqg982
Grant:
BB/W00349X/1


Publisher:
Springer Nature
Journal:
Nature Protocols More from this journal
Publication date:
2025-10-13
Acceptance date:
2025-07-29
DOI:
EISSN:
1750-2799
ISSN:
1754-2189
Pmid:
41083772


Language:
English
Keywords:
Pubs id:
2300677
Local pid:
pubs:2300677
Source identifiers:
W4415126684
Deposit date:
2026-04-27
ARK identifier:

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