2D Class Averaging Service
- Evaluating sample homogeneity and aggregation state
- Identifying conformational heterogeneity and supporting grouped modeling
- Building initial 2D models for pre-3D reconstruction comparison
- Optimizing cryo-sample preparation and improving micrograph quality
- Characterizing conformational diversity of large biomolecules such as viruses, ribosomes, and membrane complexes
2D class averaging is a critical image processing technique in single-particle cryo-electron microscopy (Cryo-EM SPA), widely used to improve the signal-to-noise ratio (SNR), clarity, and structural feature recognition of particle images. The core principle involves registering and clustering thousands of particle images, grouping those with similar orientations, and averaging them to enhance representative signals while suppressing background noise. This process ultimately produces high-resolution 2D class averages.
Compared to raw micrographs, 2D class averages provide a more accurate representation of particle conformations from different orientations, forming a solid foundation for downstream 3D reconstruction. Additionally, this step efficiently removes contaminated or incomplete particles, improving overall data quality and enabling more reliable structural analysis and sample assessment.
Figure 1. 2D Class Averaging and 3D Reconstruction
MtoZ Biolabs offers high-quality 2D Class Averaging Service that combines high-throughput image processing pipelines and advanced algorithm platforms to help clients obtain clear and reliable 2D average images, accelerating 3D reconstruction and molecular mechanism exploration.
Services at MtoZ Biolabs
In our 2D Class Averaging Service, we provide end-to-end support from image acquisition to data processing, including:
· Raw image quality assessment and automatic particle picking
· Multi-round 2D clustering and alignment optimization
· Conformational state identification and classification
· Display and annotation of 2D class average images
· Standardized data export compatible with cryoSPARC, RELION, and other platforms
We support both negative staining and cryo-EM image sources, covering sample characterization needs across different research stages.
Why Choose MtoZ Biolabs?
The 2D Class Averaging Service at MtoZ Biolabs is built upon a high-performance computing platform and advanced image analysis algorithms, offering remarkable benefits in data quality and processing efficiency:
✅ Enhanced Signal-to-Noise Performance: Efficient alignment and clustering algorithms significantly improve image quality from low-contrast raw data.
✅ Conformation Heterogeneity Compatibility: Effectively classifies heterogeneous particles to support structural polymorphism analysis.
✅ Fully Standardized Workflow: Implements a standardized image processing pipeline to ensure reproducibility and publication-ready results.
✅ Broad Platform Compatibility: Seamlessly integrates with 3D reconstruction and modeling workflows, supporting publication and patent submission.
✅ One-Time-Charge: Our pricing is transparent, no hidden fees or additional costs.
Applications
Our 2D Class Averaging Service is widely used in structural biology, vaccine development, and protein engineering, suitable for:
Case Study
Case 1: 2D Class Averaging Supports Conformational Polymorphism Analysis of the Mammalian Retromer Complex
In a study of the mammalian retromer complex, researchers utilized single-particle cryo-EM and 2D class averaging to reveal multiple assembly states during endosomal transport. Through extensive 2D classification, they identified various conformations of retromer, including heterotrimers, dimers, tetramers, and linear polymers, all exhibiting significant flexibility. These representative 2D class averages laid the groundwork for 3D reconstruction and critical interface mapping, advancing the understanding of retromer as a dynamic scaffold involved in endosomal sorting pathways. This study demonstrates the essential role of 2D class averaging in uncovering conformational polymorphism in heterogeneous complexes.
Kendall, A. K. et al. Structure. 2020.
MtoZ Biolabs offers high-quality 2D Class Averaging Service for conformational screening and early-stage structural modeling of protein complexes, facilitating the efficient progress of cryo-EM structural determination projects.
FAQ
Q1: How can I determine if my sample is suitable for 2D class averaging?
2D class averaging is ideal for datasets containing tens of thousands of particles with diverse orientations. MtoZ Biolabs provides sample evaluation services to assess data quality and optimize analysis strategies.
Q2: What structural features can be identified through 2D class averaging?
By averaging multiple similar particles, 2D class averaging enhances the SNR and reveals structural features such as overall morphology, symmetry, flexible regions, and conformational variability, aiding subsequent 3D modeling.
Q3: Do you support 2D class averaging from negative-stain images?
Yes. Although lower in resolution, negative-stain EM images can still be used to assess particle shape and aggregation. MtoZ Biolabs has extensive experience in processing negative-stain images for early-stage screening.
MtoZ Biolabs is committed to providing robust image processing solutions for structural biology. Our 2D Class Averaging Service meets diverse particle analysis needs, helping researchers enhance data quality and streamline 3D reconstruction workflows. For more information, please contact us for a free consultation.
How to order?
