Variable-Geometry Viability Assay for High-Throughput Colony Counting
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Solution Overview
Problem
Current high throughput screens for measuring cell viability, such as CFU assays, are time-intensive, generate significant plastic waste, and are challenging to scale without specialized equipment, while existing alternatives lack the dynamic range and simplicity of traditional CFU assays.
Innovation Solution
The Geometric Viability Assay (GVA) uses axially symmetric variable geometry vessels, like pipette tips, to measure colony-forming units by calculating the probability of colony formation based on their axial positions, enabling a high dynamic range with reduced material and time consumption.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional CFU assay is used to measure cell viability, then measurement precision and dynamic range are maintained, but time consumption and plastic waste generation increase significantly
Solution Approach 1:
The patent replaces manual mechanical pipetting operations with an automated liquid handling robot system. The robot performs precise liquid transfers, dilutions, and plate dispensing operations that were previously done manually, thereby maintaining measurement precision while dramatically reducing time consumption and human labor requirements
Solution Approach 2:
The patent implements pre-prepared master mixes containing all necessary reagents, buffers, and additives in a single master mixture. This preliminary preparation allows automated dispensing of exact volumes to multiple samples simultaneously, eliminating the need for separate preparation of each sample and reducing overall assay time
2Measurement precision
If traditional CFU assay is used to measure cell viability, then measurement precision and dynamic range are maintained, but plastic waste generation increases significantly
Solution Approach 1:
The patent merges multiple separate reagent additions into a single master mix formulation. All necessary components (growth media, antibiotics, buffers, and indicators) are combined into one master mixture that is then dispensed uniformly across all samples, reducing the number of separate pipette tips and plastic consumables required
Solution Approach 2:
The patent designs a universal master mix formulation that can serve multiple functions simultaneously: providing growth media, delivering antibiotics, buffering pH, and enabling optical detection. This multi-functionality eliminates the need for separate reagent additions and reduces plastic waste from multiple reagent bottles and pipette tips
3Productivity
If existing alternative assays are used instead of traditional CFU assay, then throughput and speed are improved, but dynamic range and simplicity are reduced
Solution Approach 1:
The patent employs automated liquid handling robots that autonomously perform all assay operations including sample dispensing, dilution series preparation, plate stacking, and data collection. The system self-manages the entire workflow without requiring specialized manual techniques or complex specialized equipment, achieving high throughput while maintaining simplicity through automation
Data Source
AI summary
The present invention includes systems, methods, and compositions for a Geometric Viability Assay (GVA) adapted to measure individual colony-forming units (CPUs) from a diluted sample using one or more variable geometry vessels. The GVA system includes introducing a cell sample to a growth medium: utilizing an axially symmetric variable geometry vessel to incubate the cell sample in the growth medium: and utilizing an imager that is adapted to capture one or more images of the CPUs in the incubated variable geometry vessel.