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WORC-Flow Quest™

3Rs Quest Library

NC3Rs DRIVER Recommendations

Test practical knowledge of the six DRIVER recommendations for rigorous design and transparent reporting of in vitro experiments.

Developed by an international working group of experts, the DRIVER recommendations set out six items – the core principles that support researchers design high-quality in vitro studies and report them transparently. The recommendations also include the essential information required by reviewers and editors to assess the methodological rigour, reliability and reproducibility of the work.

Read the original NC3Rs DRIVER recommendations ↗

Source acknowledgement: Educational summaries on this page are adapted and paraphrased from the NC3Rs DRIVER recommendations. Use the linked NC3Rs pages for the original authoritative wording and interactive content.

Learn first, then test yourself. Each section below gives a more detailed overview of the DRIVER recommendation and links directly to the NC3Rs interactive resource and worked examples. When ready, start the existing 15-question WORC-Flow Quest™.

6 Quests • 90 questions
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DRIVER Recommendation 01

Experimental unit

Source: NC3Rs DRIVER recommendations

Correctly identifying the experimental unit is fundamental to understanding what is genuinely replicated, what the sample size really is and what conclusions the experiment can support. DRIVER distinguishes the experimental unit (independently assigned to an intervention), the biological unit (the entity about which an inference is made) and the observational unit (where measurements are taken).Adapted and paraphrased from NC3Rs DRIVER recommendations.

What to understand before the Quest

Source: NC3Rs DRIVER recommendations
  • Independent experimental units can be assigned and treated independently and should not influence one another.
  • Genuine replication increases the number of independent experimental units; repeated measurements from one unit do not.
  • Sub-sampling can improve precision, but treating sub-samples as independent creates pseudoreplication and can distort statistical inference.
  • Increasing experimental units strengthens evidence within the sampled population; increasing biological units can improve generalisability.
  • DRIVER recommends explicitly reporting experimental and biological units rather than relying on ambiguous “biological/technical replicate” terminology.
Adapted and paraphrased from NC3Rs DRIVER recommendations.
Design focusIdentify experimental + biological units before the experiment.
Reporting focusDefine the units and explain how sub-sampling was handled.
15 questionsWorked examplesPseudoreplicationGeneralisability
📘 1: Read the NC3Rs Detailed recommendation ↗🔎 2: Take the NC3RS Interactive guide and get a Great Understanding ↗
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DRIVER Recommendation 02

Risk of bias

Source: NC3Rs DRIVER recommendations

Bias can systematically shift experimental results away from the underlying truth. DRIVER asks researchers to identify where bias could enter an in vitro experiment—from allocation and handling through outcome assessment and analysis—and choose proportionate ways to reduce it.Adapted and paraphrased from NC3Rs DRIVER recommendations.

What to understand before the Quest

Source: NC3Rs DRIVER recommendations
  • Allocation bias can arise when experimental units are assigned to groups in ways that create systematic differences.
  • Performance bias can arise from unequal handling, timing or environmental conditions across groups.
  • Detection bias can arise during measurement or assessment, particularly where judgement or order effects matter.
  • Mitigations include true randomisation, masking/blinding, balanced conditions, randomised or alternating assessment order and fit-for-purpose measurement methods.
  • Reporting should state exactly how mitigation was implemented—and justify situations where it was impractical or inappropriate.
Adapted and paraphrased from NC3Rs DRIVER recommendations.
Design focusIdentify sources of bias and select mitigation strategies.
Reporting focusDescribe what was done, how it was done and any justified limitations.
15 questionsRandomisationMaskingAllocation • performance • detection
📘 1: Read the NC3Rs Detailed recommendation ↗🔎 2: Take the NC3RS Interactive guide and get a Great Understanding ↗
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DRIVER Recommendation 03

Experimental model

Source: NC3Rs DRIVER recommendations

The experimental model must be capable of answering the biological question. DRIVER covers model selection, maintenance, characterisation and quality control—from relatively simple cell-free systems and cell cultures to co-cultures, organoids and organ-on-chip or microfluidic models.Adapted and paraphrased from NC3Rs DRIVER recommendations.

What to understand before the Quest

Source: NC3Rs DRIVER recommendations
  • Choose a model around the biology the research question actually requires, while considering generalisability, feasibility and maintenance needs.
  • Define and control important culture and microenvironmental conditions such as temperature, CO₂, humidity, media and substrate.
  • Authentication, contamination checks, phenotypic/genotypic stability and other quality controls support confidence in the model.
  • Novel or adapted models should be characterised and, where appropriate, benchmarked against existing models.
  • Report model identity, source, relevant donor/sample characteristics, maintenance conditions, experimental conditions and validation/QC information.
Adapted and paraphrased from NC3Rs DRIVER recommendations.
Design focusSelect a fit-for-question model and establish maintenance + QC.
Reporting focusDescribe the model and conditions sufficiently for interpretation and replication.
15 questionsOrganoidsOrgan-on-chipAuthentication & QC
📘 1: Read the NC3Rs Detailed recommendation ↗🔎 2: Take the NC3RS Interactive guide and get a Great Understanding ↗
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DRIVER Recommendation 04

Experimental procedures

Source: NC3Rs DRIVER recommendations

Experimental procedures cover what was done, when and how often, and the equipment, reagents, software and materials used. DRIVER emphasises planning and testing procedures before experiments begin and documenting enough detail for others to verify and reproduce the work.Adapted and paraphrased from NC3Rs DRIVER recommendations.

What to understand before the Quest

Source: NC3Rs DRIVER recommendations
  • Record reagents, preparation, quantities/concentrations, equipment make/model/calibration, timings, step-by-step methods and software versions/settings.
  • Document deviations from established or published protocols rather than relying on a citation alone.
  • Use quality control routinely; perform fit-for-purpose assessment when methods, reagents or equipment are introduced or changed.
  • Calibration should occur at suitable intervals, and new biological reagent lots may require checks for lot-to-lot variability.
  • Detailed protocols can be placed in stable protocol repositories; identifiers such as RRIDs and suitable chemical identifiers can improve unambiguous reporting.
Adapted and paraphrased from NC3Rs DRIVER recommendations.
Design focusDocument procedures and test that components are fit for purpose.
Reporting focusReport what, when, how often, and exactly which equipment/reagents were used.
15 questionsProtocolsCalibrationReagents & equipment
📘 1: Read the NC3Rs Detailed recommendation ↗🔎 2: Take the NC3RS Interactive guide and get a Great Understanding ↗
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DRIVER Recommendation 05

Experimental groups and exclusions

Source: NC3Rs DRIVER recommendations

Excluding data can sometimes be necessary, but selective or subjective exclusions can alter interpretation and introduce bias. DRIVER therefore emphasises anticipating exclusions, defining objective criteria before outcomes are known, applying them consistently and reporting all exclusions transparently.Adapted and paraphrased from NC3Rs DRIVER recommendations.

What to understand before the Quest

Source: NC3Rs DRIVER recommendations
  • Potential exclusions include technical failures, experimental criteria and analysis criteria.
  • Where possible, inclusion/exclusion thresholds should be objective, predefined and applied equally across groups.
  • Analysis-related exclusions should be specified in advance; some technical failures cannot realistically be predicted.
  • Documenting criteria in an experimental plan reduces the risk of decisions being influenced by observed outcomes; preregistration can add transparency where appropriate.
  • Report data from all experimental groups and controls and clearly describe and justify exclusions.
Adapted and paraphrased from NC3Rs DRIVER recommendations.
Design focusPlan how all data will be handled and predefine criteria where possible.
Reporting focusReport all groups and controls and transparently justify exclusions.
15 questionsExclusion criteriaControlsSelective reporting
📘 1: Read the NC3Rs Detailed recommendation ↗🔎 2: Take the NC3RS Interactive guide and get a Great Understanding ↗
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DRIVER Recommendation 06

Data availability and presentation

Source: NC3Rs DRIVER recommendations

How data are managed, shared and visualised affects whether results can be understood, checked and reused. DRIVER recommends planning data management and presentation during experimental design rather than treating them as publication-stage decisions.Adapted and paraphrased from NC3Rs DRIVER recommendations.

What to understand before the Quest

Source: NC3Rs DRIVER recommendations
  • Plan storage, metadata, repositories, access conditions and licensing early, while respecting ethical, legal and commercial constraints.
  • Apply FAIR principles so data are Findable, Accessible, Interoperable and Reusable.
  • Plotting should reflect the true experimental unit; displaying sub-samples as though they were independent can visually imply pseudoreplication.
  • Choose visualisations that communicate individual values or the distribution and variability of the data, depending on dataset size.
  • Document image processing and retain originals; report representative images and manipulations transparently.
Adapted and paraphrased from NC3Rs DRIVER recommendations.
Design focusPlan transparent data management, sharing and visualisation.
Reporting focusMake data verifiable/reusable and present distributions and study design clearly.
15 questionsFAIR dataVisualisationMetadata & repositories
📘 1: Read the NC3Rs Detailed recommendation ↗🔎 2: Take the NC3RS Interactive guide and get a Great Understanding ↗

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