Pollen Extraction via Cryogenic Freezing and Image Analysis
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Solution Overview
Problem
Current methods for quantifying pollen are inefficient, inaccurate, and time-consuming due to the microscopic size and light weight of pollen, making it difficult to capture and count, especially in plants like corn where pollen is produced in large volumes and shed at different times, leading to challenges in measuring pollen production per tassel and evaluating genetic and environmental effects.
Innovation Solution
A method involving blending the plant with a liquid to separate pollen, followed by filtration and image analysis using software to count pollen grains, allowing for high-throughput and accurate quantification of pollen production, which can be applied to both pre-shed and post-shed pollen.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If pollen is captured using sticky traps or clear plastic bags in the field, then pollen can be collected for analysis, but not all pollen is captured accurately and stress effects are imposed on the tassel
Solution Approach 1:
The patent applies preliminary action by collecting tassels before pollen shed occurs and performing extraction in a controlled laboratory environment. This prevents stress effects that would occur with field-based collection methods while ensuring all pollen is captured. The tassels are harvested intact and processed offline, eliminating the need for sticky traps or plastic bags that impose stress on living plants.
Solution Approach 2:
The patent introduces liquid nitrogen as an intermediary substance to freeze and preserve pollen within the tassel structure. This allows complete pollen capture without physical disruption or stress to the tassel. The liquid nitrogen penetrates the tassel and freezes pollen in place, enabling accurate quantification while maintaining tassel integrity during collection and transport.
2Ease of manufacture
If pre-shed pollen is extracted using dry grinding or chemical methods, then pollen can be separated from the plant, but the process is time-consuming and may damage the pollen
Solution Approach 1:
The patent fundamentally changes the physical state parameter by using liquid nitrogen to freeze pollen, making it brittle and easy to separate from the tassel structure. This phase change from liquid to solid allows rapid pollen release through simple mechanical disruption, eliminating time-consuming chemical treatments and significantly reducing extraction time while preserving pollen integrity.
3Measurement precision
If pollen counting is performed using visual estimation or manual methods, then pollen production can be evaluated, but the process is time-consuming and lacks accuracy for large volumes of pollen
Solution Approach 1:
The patent replaces manual visual estimation and mechanical counting methods with automated image analysis technology. Digital images of the extracted pollen are processed by computer algorithms that automatically count and measure pollen grains, providing high-precision measurements for large volumes without time constraints. This substitution of mechanical/manual systems with automated optical systems simultaneously improves accuracy and eliminates counting time limitations.
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 approach enables quick, reliable, and reproducible pollen quantification, facilitating the evaluation of pollen production for plant breeding, genetic advancement, and commercial production by providing statistically accurate and efficient pollen counts per tassel, even for large numbers of samples.
Implementation Method 1
The tassel is placed in liquid nitrogen to freeze the pollen
Implementation Method 2
The frozen tassel is blended with liquid using a blender
Implementation Method 3
Pollen is extracted from the liquid/plant suspension through a filter or strainer sized to pass relevant pollen grains
Data Source
AI summary
An apparatus and method of quantitatively obtaining a measurement of pollen of a plant. One method of counting comprises imaging the sample with the pollen well-distributed in the focal plane of the imager. Image evaluation software can identify and count objects in the image that are consistent with pollen. Total pollen count for the plant can be derived from the count of pollen of the sample, proportionality of the sample volume to the starting volume, and proportionality of area of sample imaged to total area of sample. Pollen quantification can be used for research or commercial production decisions relative to the plant or its seed.