> For the complete documentation index, see [llms.txt](https://guide.cryosparc.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-reconstruction/job-homogeneous-ab-initio-refinement-beta.md). # Job: Homogeneous Ab-Initio Refinement (BETA) ## At a Glance Use CryoSPARC's *ab-initio* algorithm (based on stochastic gradient descent) to refine a dataset to high resolution from scratch, while preserving gold standard independence of two half-sets and half-maps. ## Description {% hint style="warning" %} Homogeneous Ab-Initio Refinement should generally be tried only once traditional global refinements fail to produce reasonable results. See Recommended Alternatives for more information. {% endhint %} In [some challenging cases](https://www.biorxiv.org/content/10.1101/2025.09.08.674935v1), refinement of a 3D map to high resolution using CryoSPARC's [gradient descent](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-reconstruction/job-ab-initio-reconstruction#stochastic-gradient-descent) algorithm for *ab-initio* reconstruction outperforms the traditional [expectation maximization algorithm](https://guide.cryosparc.com/expectation-maximization-in-cryo-em) used in refinement jobs \[1]. However, directly using the [Ab-Initio Reconstruction](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-reconstruction/job-ab-initio-reconstruction) job to produce high-resolution volumes makes resolution estimation and validation difficult, since Ab-Initio Reconstruction does not use gold standard half-sets. **New in CryoSPARC v5.0,** This job splits input particles into two half-sets and refines independent half-maps each using the same process as [Ab-Initio Reconstruction](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-reconstruction/job-ab-initio-reconstruction). Notably, this job is still *ab-initio* in that it does not use any input volume; both half maps are reconstructed independently from scratch from the data alone, and kept aligned to each other by a 3D map alignment step within the job. This job can only produce a single homogeneous output rather than multiple classes. The output half-maps can be further refined downstream using [Local Refinement](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/local-refinement/job-new-local-refinement-beta) which can sometimes further improve map quality. Note that using a global refinement downstream, like [Homogeneous](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-refinement/job-homogeneous-refinement) or [Non-Uniform Refinement](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-refinement/job-non-uniform-refinement-new), typically will not yield results superior to using standard Ab-initio Reconstruction followed by refinement. ## Inputs ### Particle stacks Particles from which half maps will be produced. ## Commonly Adjusted Parameters See [Ab-Initio Reconstruction](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-reconstruction/job-ab-initio-reconstruction#commonly-adjusted-parameters) for parameter information. ## Outputs ### Particles Particles with poses and half-set splits used to produce the half maps. ### Volumes Unlike other refinement jobs in CryoSPARC, Homogeneous Ab-Initio Refinement produces only half maps. ### Unused particles Particles which were not used. This typically means that the refinement converged before all particles had been aligned. If you wish to align all particles, set `Num particles to use` to a value greater than or equal to the number of input particles. ## Common Next Steps To produce an averaged, filtered full map you can either use [Homogeneous Reconstruction Only](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-refinement/job-homogeneous-reconstruction-only) with the particles output, or use the particles and half maps in [Local Refinement](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/local-refinement/job-new-local-refinement-beta) with `Initialize from input half-maps` turned on. ## Recommended Alternatives Homogeneous Ab-Initio Refinement currently requires far more iterations than other techniques to produce a final map. For this reason, this job should only be used when both: * Particle images produce high-quality 2D classes, indicating that real signal is present in the particle stack * Global refinements of particles (e.g., [Homogeneous](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-refinement/job-homogeneous-refinement) or [Non-Uniform Refinement](https://guide.cryosparc.com/processing-data/all-job-types-in-cryosparc/3d-refinement/job-non-uniform-refinement-new)) produce maps with a quality much worse than expected based on 2D Classification results ## References 1. Kookjoo Kim, Huan Li, Oliver B. Clarke (2025). High-resolution ab initio reconstruction enables cryo-EM structure determination of small particles. bioRxiv 2025.09.08.674935; doi: