AEGIS: Assay-Aware Protocol Validation and Runtime Monitoring for Open-Source Liquid Handling Robots
Authors: Priyanka V. Setty, Arvind Ramanathan, Ian Foster, Rick Stevens
Organizations: a Data Science and Learning Division, Argonne National Laboratory, Lemont, IL 60439, USA. · b Department of Computer Science, University of Chicago, Chicago, IL 60637, USA.
Abstract
Self-driving laboratories increasingly rely on low-cost liquid handlers such as the Opentrons OT-2, which ship without the pressure-based aspiration monitoring of Hamilton or Tecan systems and are typically run open-loop. Two failure modes go undetected: protocols that are syntactically valid but violate assay-specific invariants (e.g., tip reuse between a PCR template and a no-template control), and physical execution failures (partial dispense, air bubbles, missing tips) at runtime. We present AEGIS, a two-layer guardian for both. Layer 1 pairs a curated machine-readable assay rule database with an LLM that reasons over OT-2 Python code, reaching an adjusted F1 of 0.97 on a 24-protocol benchmark across five assay families and beating rules-only and LLM-only ablations across five backends; a free open-weight model ties the best proprietary one, so no paid API is required. Layer 2 fits a PCA world model to YOLO-cropped four-frame pipette trajectories; under a leakage-free leave-one-plate-out evaluation it reaches average precision 0.89 and operating-point F1 0.71 (AUROC 0.80), a deployment-faithful number that matches the live demonstration, and we characterize the small-pipette (p20) resolution limit (F1 0.47). A live demonstration on a physical OT-2 (five replicates per condition) catches planted no-tip failures deterministically and partial dispense on coloured dyes, with an always-VLM self-vote gate lifting partial-dispense recall to 5/5; transparent water is a principled limit of any front-view-only monitor, which AEGIS surfaces as low-confidence VLM reasoning rather than a wrong verdict. Cascade triage holds VLM cost near 1.63perplateversus10.33 for an always-VLM baseline. AEGIS is open source and, to our knowledge, the first system to unify pre-flight assay-aware validation with runtime visual monitoring for an open-source liquid handler.
Laboratory automation accelerates discovery, yet its adoption is constrained by the high cost, proprietary design, and limited remote supervisability of commercial liquid-handling systems. This work presents RAINBOT\textsuperscript{TM}, a low-cost, openly reproducible liquid-handling robot built by converting a consumer-grade Cartesian 3D printer (Elegoo Neptune 4 Max). The printer extruder is replaced by a precision single-channel pipette actuated through the printer's own G-code-driven X--Y--Z gantry, with plunger and tip-eject motions effected by two compact linear actuators under Python control. To make experiments transparent and remotely supervisable, a browser-based digital twin is implemented to synchronise bidirectionally with the physical platform, mirroring kinematics and pipetting states in real time and exposing remote monitoring, intervention, and an emergency stop from any web browser. As a proof of concept, RAINBOT\textsuperscript{TM} performed sequential exchanges of differently coloured aqueous solutions while an integrated colour sensor quantified the resulting mixtures; measured red, yellow, and blue (RYB) responses agreed with expected mixing behaviour to within a mean absolute error of two percentage points, validating correct execution and real-time tracking. Closing the loop, the platform is coupled to the CEID\textsuperscript{TM} (Cooperative Explorer for Inverse Design) framework, which recasts experimentation from iterative manual guessing into a goal-directed inverse-design search while keeping a human in the loop. The complete hardware costs under US$1300, which is roughly an order of magnitude below entry-level commercial handlers, thereby establishing an accessible physical--virtual framework for self-driving laboratory automation.
Autonomous wet-lab experimentation requires more than plausible protocol text: biological intent, quantitative procedures, device constraints and experimental feedback must remain aligned from protocol and SOP design to code and physical execution. We developed ProtoPilot, a self-evolving multi-agent system, together with an expert-grounded benchmark and evaluation framework for testing this conversion as an experimental automation problem. The framework spans 294 synthetic-biology and molecular-biology tasks derived from 98 gold-standard protocols, wet-lab expert rubrics, device-level validity gates and real experimental tests. ProtoPilot incorporates layer-wise verifiability, multi-agent orchestration and a runtime-updated skill library to generate protocols, expand SOPs, synthesize SDK-compliant code and revise workflows from wet-lab feedback. It achieved a Top@3 expert-preference rate of 90.2%, an overall protocol-to-code gate pass rate of 89.5% and an Opentrons pass rate of 88.24%, compared with 32.35% for OpenTrons-AI. Wet-lab validation produced interpretable readouts, Sanger-confirmed products and feedback-corrected PCA-assembled DNA targets, establishing a verifiable route to autonomous experimentation. Together, these results show that the evaluation framework captures execution-relevant requirements for autonomous wet-lab automation, and that ProtoPilot can meet them by converting protocol and code generation into validated execution and feedback-guided revision.
Biological experiment protocols are written in natural language, whereas automation systems rely on predefined control commands, creating a semantic gap that limits autonomous execution. Microplate-based automatic experiments are particularly challenging due to the need to simultaneously control well mapping, sample-reagent combinations, replicate placement, and parallel dispensing. This study proposes an agent-based protocol translation framework that converts natural-language microplate-based protocols into executable control commands for a robotic laboratory platform. A Parser Agent formalizes the natural-language protocol into a structured representation, and a rule-based mapping engine deterministically incorporates the operational constraints of the robotic laboratory platform to generate device-level control commands. A heterogeneous LLM Validation Agent verifies completeness, parameter accuracy, and execution order, and triggers a self-correction loop with structured feedback when errors are detected. A sweep involving 7 Parsers and 3 Validators on randomly selected ELISA protocols evaluates how model scale and Validator type affect translation accuracy and pass rates under cross-model verification. The accuracy-latency trade-off is further verified by comparing the rule-based mapping of the proposed framework with LLM end-to-end direct mapping. Finally, Bradford assay-based protein quantification using a microplate was demonstrated on a robotic laboratory platform, validating end-to-end autonomous execution from natural-language protocols to real-world experiments. The proposed framework provides a flexible approach to narrowing the semantic gap between natural-language protocols and microplate-based self-driving laboratories.