Seed Tissue Sampling Using Cookie Cutter Punch

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Solution Overview

Problem

Current methods for sampling seed tissue, particularly for genotyping, face challenges such as inefficiency, poor reproducibility, and loss of germinative capacity, with existing techniques like drilling and automation systems being cumbersome, costly, and prone to cross-contamination.

Innovation Solution

A manual or semi-automatic method using a cookie cutter to extract a predetermined volume of seed tissue from the endosperm or cotyledon, allowing for precise control and preservation of germinative capacity, with the option to soak seeds in water for softening and using a multi-well plate for analysis.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If drilling is used to obtain powder from seed, then sample quantity can be obtained, but the process is long and tedious and leads to heating of the sample

Engineering Contradiction:
Improvesample quantityVSAvoidprocess time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

The invention extracts only the necessary portion of seed tissue using a punch tool to obtain a core sample of predetermined volume, rather than processing the entire seed through drilling. This extraction approach provides sufficient material for DNA extraction while dramatically reducing processing time and avoiding heating issues associated with extensive drilling operations.

Inventive Principle:
Principle #2Taking out (Extraction)

2Productivity

If automation systems are used for sampling, then productivity increases, but device complexity and cost increase significantly

Engineering Contradiction:
Improvesampling efficiencyVSAvoidsystem complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The punch tool is designed to be self-contained and manually operable, requiring no complex automated systems. The tool itself performs the sampling function through simple manual insertion and extraction, making the process accessible without expensive automation infrastructure while maintaining consistent sampling capability.

Inventive Principle:
Principle #25Self-service

3Reliability

If manual cutting pliers are used to take grain samples, then seed viability is preserved, but the quantity of material taken is relatively large and cannot be precisely calibrated

Engineering Contradiction:
Improveseed viabilityVSAvoidsample quantity precision
Core Design Contradiction:
ReliabilityVSManufacturing precision

Solution Approach 1:

The punch tool is designed with specific dimensional parameters (diameter and length) that define the volume of the core sample. By controlling these geometric parameters, the invention achieves precise calibration of sample quantity (e.g., 0.5-2 mg) while maintaining seed viability through minimal tissue removal compared to cutting pliers methods.

Inventive Principle:
Principle #35Parameter changes

4Productivity

If automated leveling cutting is used on continuously fed seeds, then productivity increases, but germination capacity is significantly reduced

Engineering Contradiction:
Improvesampling throughputVSAvoidgermination capacity
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The invention applies partial action by removing only a small core portion of the seed rather than leveling or cutting large portions. This minimal extraction approach obtains sufficient material for analysis while preserving the majority of the seed structure needed for germination, achieving a balance between sampling efficiency and seed viability.

Inventive Principle:
Principle #16Partial or excessive action

5Shape

If abrasive leveling is used on seeds, then sample surface is prepared, but cross-contamination risks remain and DNA extraction becomes laborious

Engineering Contradiction:
Improvesample surfaceVSAvoidcross-contamination risk
Core Design Contradiction:
ShapeVSObject-affected harmful factors

Solution Approach 1:

The punch tool extracts a discrete core sample that is physically separated from the seed surface, eliminating the need for abrasive leveling that creates dust and contamination risks. The extracted core can be directly transferred to extraction tubes, reducing handling steps and cross-contamination opportunities while providing sufficient material for efficient DNA extraction.

Inventive Principle:
Principle #2Taking out (Extraction)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This method ensures reproducible, controlled, and standardized sampling, maintaining seed viability, and allows for precise calibration of sample quantity and quality, facilitating genotyping and other analyses while ensuring traceability and efficient DNA extraction.

Implementation Method 1

using a cookie cutter designed to cut, take away, then release a determined volume of tissue

Methodology Applied
Scientific EffectMechanical cutting: Mechanical Force

Implementation Method 2

optionally soak the seed with water so as to soften it and allow the next step

Methodology Applied
Scientific EffectImbibition: Absorption (physical)

Data Source

PatentEP3097398B1Seed tissue sampling process
Publication Date: 2021.11.03 LIMAGRAIN EURO SA
  • EP3097398B1 patent drawingFigure 1
  • EP3097398B1 patent drawingFigure 2~3
  • EP3097398B1 patent drawingFigure 4

AI summary

Process for sampling tissue from seeds of given size, in particular from maize and sunflower, with or without preservation of the germinal nature of the seeds. Method for sampling seed tissue when the amount of the tissue sample must be controlled, for example within the context of genotyping or of any method for analyzing a constituent of the seed. The process performs a manual or semi-automatic withdrawal of a given sample volume from the seed(s), using a punch designed for cutting, taking, then releasing a given volume of tissue, this volume being determined in order to enable the analysis of at least one constituent of the seed or seeds. The sample of tissue from a single seed or from several seeds is placed in an identified and traceable well, then the analysis of said constituent is carried out on the content of the well(s).