Single Particle Cryo-EM

Cryo-EM Process Overview

Construct Design

Based upon literature review, modelling and any prior in-house experience. Will include any accessory proteins if required.

Small Scale Feasibility Studies

Both expression and purification experiments, usually screening a number of constructs with the aim to define the optimal host/construct combination. QC analysis is critical for selection.

Scale Up

The aim is to produce the target protein(s), in sufficient quality and quantity, to support grid preparation. At this stage we can perform a number of QC tests, including analysis by SEC, SDS-PAGE, FIDA, mass spectrometry, and negative stain EM.

Grid Preparation and Screening

In-house grid preparation using a Vitrobot IV allows for grid preparation using freshly purified proteins Grids are screened, and if appropriate, test data set(s) are collected to enable 2D and 3D classification.

Cryo-EM Full Data Collection

Access to facilities with state-of-the-art microscopes (Titan Krios) with our scientists running the experiments.

Data Processing and Modelling

In-house scalable processing power offered by the Amazon Web Services (AWS) cloud platform provides ability to process cryo-EM data rapidly and on-demand with data security paramount. No queuing for computational resource required

Hear from one of our experts

“We can support all stages of a single particle cryo-EM project from construct design through to refined structure. Protein quality is the key factor for structure determination by cryo-EM and often the most interesting targets are the toughest to produce. We have extensive experience in producing difficult proteins, including membrane proteins.”

Valerie Pye: Senior Principal Scientist

Why Choose Peak Proteins for your Cryo-EM Project

In house cryo-EM expertise, covering every stage of the process
Protein quality is the key factor for cryo-EM and often the cryo-EM targets are some of the toughest to produce. At Peak Proteins we have a solid track record in being able to produce challenging protein targets, including membrane proteins.
We have the tools to perform quality checks prior to cryo-EM grid preparation to ensure the best protein is taken forward (e.g., analytical SEC, MS-MS, FIDA, negative stain EM).
In house Vitrobot IV for grid preparation.
Access to state-of-the-art microscopes for grid screening and data collection (Glacios, Titan Krios) with our scientists running the experiments.
- Access to facilities at LonCEM, eBIC (Diamond Light Source) and the Universities of Manchester, Leicester and Leeds.
In-house data processing
Secure and scalable processing power offered by Amazon Web Services (AWS) cloud platform provides ability to process cryo-EM data rapidly and on-demand. No queuing for computational resource required.
Project risk management.
- A cryo-EM project can be an iterative process where individual stages need further optimisation.
- Our workflow include various stop/go points to reduce and manage this risk.
- We regularly meet with clients and will offer honest assessments of project status and advise around cost/risk.

5HT2a Cryo-EM structure solved in house.
Gene to Structure