MP - Mass Photometry
Please acknowledge the SIP core facility ( RRID: SCR_018986) in publications, on posters, or in talks if you use any instruments in the SIP core facility. Please include SIP's RRID (RRID: SCR_018986) and the grant numbers for instruments funded through instrumentation grants in your acknowledgements. This is a requirement from the funding agencies and is crucial for future funding. Find example text on theÌýÌýor the individual instrument pages.
Mass Photometry in Biochemistry, Biophysics and Structural Biology
Mass photometry (MP) is a cutting-edge technique that allows for the precise measurement of the mass of single molecules in solution. This innovative method is particularly advantageous because it enables the study of biomolecules in their native state, without the need for ionisation or fragmentation, which are typically required in traditional mass spectrometry.
The fundamental principle behind mass photometry involves the detection of light interference. When a focused beam of light strikes a glass surface, a portion of the light is directly reflected, while another portion is scattered by biomolecules as they adhere to the surface. The interaction between the reflected and scattered light produces an interferometric contrast, which is directly correlated with the molar mass of the scattering biomolecules. This unique approach allows researchers to accurately measure the mass of proteins, nucleic acids, and other biomolecules in solution.
Mass photometry offers valuable insights into the molecular weight and oligomeric states of biological macromolecules and complexes. This makes it an invaluable tool in structural biology, where understanding the assembly and structure of biomolecular complexes is crucial. Additionally, in biochemistry and biophysics, MP can be used to study the behaviour and interactions of biomolecules under various conditions, providing a deeper understanding of their functions and dynamics.
Beyond basic research, mass photometry also holds significant potential in drug discovery. By enabling the detailed analysis of biomolecular interactions and complex formation, MP can for example aid in the identification of potential drug targets. It can help in the optimisation of therapeutic molecules, or facilitate quality control of AVV samples by providing a fast and easy way for the quantification of empty, partially filled, full and overfull AAVs. ÌýOverall, mass photometry is a powerful technique that is rapidly becoming essential in the study of biological systems.
Key highlights of MP in Biology, Biochemistry, and Biophysics
- Accurate molecular weight measurementsin solution without the need for labelling or modifying the sample, preserving its native state.
- Compatible with a wide range of biomolecules, including proteins, RNA, DNA, complexes, vesicles, micelles, adeno-associated viruses (AAVs), and even some polymers.
- Detailed information on sample heterogeneity, enabling the measurement of masses and the number of molecules in different subpopulations within a sample.
- Compatible with a broad range of buffers, allowing flexibility in experimental conditions.
- Effective with low concentrations (100 pM to 100 nM) and small sample volumes (15-20 µL), making it suitable for precious or limited samples.
Ìý
Instrument and Accesories
Instrument
TwoMP mass photometer from Refeyn
- Mass range: 30 kDa – 5 MDa
- Resolution (FWHM): 25 kDa @ 66 kDa, 60 kDa @ 660 kDa)
- Mass precision: ± 2%
- Mass error: ± 5% (single measurement)
- Concentration range: 100 pM – 100 nM
- Sensitivity: << 1 ng of protein
- Wavelength: 488 nm
- Field of view: 4 x 11 μm (@ 500 Hz) up to 12 x 17 μm (@ 135 Hz)
- Pixel size: 12 nm
AccessoriesÌý
- ÌýRefeyn AcquireMP for data acquisition
- ÌýRefeyn DiscoverMP for data analysis
- ÌýSample well cassettes (6 sample wells/cassette)
- ÌýAlignment tool, tweezers, and magnetic slide holders
- ÌýLens tissue to clean the instrument objective
Ìý