How does FIDA increase Cryo-EM scan success rates?
01
Reduce your time to publication
- Key sample properties identified in just 4 minutes
- Upfront rejection of samples of suboptimal quality samples reduces costs and saves time spent on samples that are not suited for Cryo-EM of droplet formation
- Informed troubleshooting
02
Key sample properties determined
- Ensure homogeneity with polydispersity index (PDI)
- Quantify aggregation
- Measure absolute viscosity
- Detect and quantify stickiness – prior to cryo-EM scan
- Check structural integrity - thanks to absolute size (Rh) measurement
03
Directly in Cryo-EM relevant concentrations
- FIDA users do not need to dilute their samples to get key pre-cryo-EM parameters
04
Native conditions
- No buffer constraints
- No detergents, salt, pH or concentration constraints, provide increased flexibility.
- Triple temperature control
- UV-detector allows to work with unlabelled proteins.
05
Lower sample consumption
- Only hundred nanolitres per condition in the screening stage
- Down to 4 μL analyte, 40 nL indicator for characterisation
06
Boosted process efficiency
- FIDA users reduce overall cryo-EM cost by discarding unsuitable samples upfront
- FIDA users avoid wasting time and financial losses incurred by scanning samples of poor quality
- Low energy consumption of the Fida instrument
- Low sample usage (down to 4 μL analyte, 40 nL indicator)
- Full automation saves time
- Speed of QC process minimises time-related sample degradation
07
Improved structural resolution and data reproducibility
- Pre-cryo-EM in-depth sample characterization
- Flexible dilution options, native conditions
- Optimize sample deposition through enhanced knowledge of your sample
- Flexible buffer conditions allow FIDA users to work in a wide range of pH and salt concentrations
08
Automated and temperature controlled
- Autosampler fits 2 x 96 well plates or 2 x 50 vials. Save your time for what really matters.
- Keep samples stable at 5°C during the duration of an assay
09
Work with membrane proteins and nanodiscs
- Assess membrane proteins and lipid nanodiscs thanks to FIDA’s detergent compatibility.
Success of CRYO-EM assay depends on the sample quality.
FIDA users select the adequate sample upfront thanks to fast and efficient pre cryo-EM analysis.
Quantitative data about viscosity, aggregation, homogeneity and oligomeric state,
measured in one assay, allows FIDA users to select only the best samples,
and avoid the time and cost loss incurred due to scanning samples of poor quality.
On top of that, having an in-depth knowledge of the sample makes it easier to perform grid preparation.
Quantitative data about viscosity, aggregation, homogeneity and oligomeric state,
measured in one assay, allows FIDA users to select only the best samples,
and avoid the time and cost loss incurred due to scanning samples of poor quality.
On top of that, having an in-depth knowledge of the sample makes it easier to perform grid preparation.
Absolute size (Rh)
- Hydrodynamic radius measurement from 0.5 to 500 nm
- Makes it possible to ensure the basic feasibility of a sample/that the sample is within practical size limits of cryo-EM
- Globular size change (Rh) down to 5%
Aggregation
- Quantified FIDA readout is delivered as a number of aggregates in the sample
- Quantification of aggregates performed ubiquitously – all throughout the measurement
- Aggregates are presented visually as signal spikes on raw FIDA data
- No constraints on size of aggregates
Stickiness – fully visible
- Reveals sample hydrophobicity – more informed workflow helps to achieve optimal layer thickness
- Awareness of sample stickiness helps to troubleshoot issues with ice thickness and ultimately deposit a suitable monolayer of macromolecules
- FIDA is the only technology on the market that delivers stickiness readout
Sample polydispersity index
- Ensure homogeneity & spot the optimal purification stage to avoid over-purification.
- Measured in high concentrations – FIDA users have no need to dilute the sample to assess homogeneity
- Insight into potential protein denaturation and/or deformation pre-vitrification
Viscosity as an absolute measurement
- Quantified viscosity measurement – allows FIDA users to assess suitability of the sample to cryo-scanning and customise sample deposition settings.
01
FIDA may also be a useful tool in prevention of denaturation issues
Since denaturation changes the globular size of a molecule, FIDA’s absolute size measurement can indicate those changes in samples pre-vitrification.
This ensures that samples that denature pre-vitrification are excluded.
This ensures that samples that denature pre-vitrification are excluded.
02
FIDA may also be a useful tool in prevention of denaturation issues
Secondly, FIDA can be used for buffer screening. FIDA’s buffer flexibility makes it possible to test the behaviour of the molecule in buffers with diverse pH or salt concentration.
Since buffers impact the stability of the protein (due to the buffer-related surface charge), altering buffer composition changes the molecule's behaviour upon freezing. Using FIDA to choose the right buffer can help to prevent denaturation.
Since buffers impact the stability of the protein (due to the buffer-related surface charge), altering buffer composition changes the molecule's behaviour upon freezing. Using FIDA to choose the right buffer can help to prevent denaturation.
03
FIDA may also be a useful tool in prevention of denaturation issues
On top of that, since FIDA does not require sample delusion, a sample is QCed at the same concentration as the sample used for vitrification, increasing the applicability of QC results obtained.
04
FIDA may also be a useful tool in prevention of denaturation issues
Moreover, FIDA’s triple temperature control makes it possible to maintain stable temperature during the quality check.
Benefits
Sample QC at the concentration used during vitrification
Identification of samples with a high likelihood of CRYO-EM success
Sample QC at the concentration used during vitrification
Direct impact on success rate and speed-to-publication
A QC assay in 4 minutes
Stability control
Informed troubleshooting
Features
Undiluted measurement
No buffer restrictions
Quantified aggregation
Polydispersity index
PDB structure correlator
Just 120 nL of sample consumed for triplicate measurement
Samples can be analysed and stored at desired temperatures
Recoverable sample – can be used directly for vitrification
Measures
Accurate determination of dilute phase concentration
Relative droplet size distributions
Kinetics of droplet formation and maturation into amyloid fibrils
Determination of binding affinity between the polypeptide undergoing LLPS and LLPS-modulating compounds
Become a user
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