Sperm Nuclei Calibration via Sonication and Dye

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Solution Overview

Problem

The production of sperm nuclei for flow cytometer calibration is hindered by centralized manufacturing requirements, regulatory issues, use of hazardous chemicals, lengthy manufacturing processes, and suboptimal signal generation, leading to decreased productivity in sperm cell analysis and sorting.

Innovation Solution

A method involving unsorted sperm nuclei combined with a DNA selective dye and an aggregation-reducing compound, sonicated to create optimally stained sperm nuclei for rapid and safe calibration of flow cytometers, eliminating the need for hazardous preservatives and reducing manufacturing time.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Duration of action of stationary object

If sperm nuclei are manufactured using conventional centralized methods, then long-term storage and preservation are achieved, but hazardous chemicals such as azide are required and regulatory issues arise

Engineering Contradiction:

Improvestorage durationVSAvoidhazardous chemicals

Core Design Contradiction:

Duration of action of stationary objectVSObject-affected harmful factors

Solution Approach 1:

The patent changes the chemical composition parameters of the storage medium by replacing hazardous chemicals like azide with non-hazardous alternatives such as egg yolk-based buffers and commercial sperm extenders. This parameter change maintains long-term storage capability while eliminating harmful factors.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent employs disposable, non-hazardous storage buffers that can be easily discarded after use, eliminating the need for hazardous preservatives. These short-living storage solutions are designed for single use or limited storage periods without requiring dangerous chemicals.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

2Quantity of substance

If sperm nuclei are manufactured by pooling samples from multiple animals, then adequate quantity is achieved, but manufacturing complexity and regulatory issues increase

Engineering Contradiction:

Improvesperm nuclei quantityVSAvoidmanufacturing process complexity

Core Design Contradiction:

Quantity of substanceVSDevice complexity

Solution Approach 1:

The patent segments the manufacturing process into simple, discrete steps that can be performed independently. Instead of complex pooling from multiple animals, the method uses straightforward processing of single-animal samples through sequential operations: staining, sonication, and storage.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent inverts the conventional approach by using unsorted, whole sperm samples directly from a single animal rather than pooling and sorting from multiple animals. This inversion simplifies the manufacturing process while still achieving adequate quantity for calibration.

Inventive Principle:

Principle #13The other way round (Inversion)

3Productivity

If conventional sperm nuclei manufacturing methods are used, then nuclei are produced, but signal generation by flow cytometer is suboptimal and calibration time is lengthy

Engineering Contradiction:

Improvecalibration speedVSAvoidsignal generation quality

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The patent applies preliminary staining of sperm nuclei with DNA-selective dyes before flow cytometer calibration. This preliminary action ensures optimal signal generation is achieved in advance, eliminating the need for lengthy calibration procedures and improving both speed and quality.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent uses sonication (mechanical vibration) to process the sperm nuclei, which improves their physical properties and enhances signal generation. The vibration-based processing optimizes the nuclei for flow cytometer detection, achieving both rapid calibration and high signal quality.

Inventive Principle:

Principle #18Mechanical vibration

4Measurement precision

If sperm nuclei are sonicated with DNA selective dye, then signal generation is improved, but manufacturing time and process complexity increase

Engineering Contradiction:

Improvesignal generation qualityVSAvoidmanufacturing time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent merges multiple functions into a single integrated process: the DNA-selective dye staining and sonication are combined into one simultaneous operation. This merging achieves optimal signal generation while minimizing manufacturing time, as both functions are performed together rather than sequentially.

Inventive Principle:

Principle #5Merging (Combining)

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 enables rapid, safe, and efficient production of improved sperm nuclei for flow cytometer calibration, enhancing productivity and reducing regulatory concerns by eliminating the use of hazardous chemicals and simplifying the manufacturing process while achieving higher quality signal generation.

Implementation Method 1

combining a DNA selective dye with the unsorted sperm cell sample to create a sperm cell mixture

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Implementation Method 2

sonicating the sperm cell mixture to create stained sperm nuclei

Methodology Applied

Scientific EffectUltrasonic Vibration: Ultrasonic Vibration

Data Source

PatentUS20220339298A1Sperm nuclei and methods of their manufacture and use

Publication Date: 2022.10.27 INGURAN LLC

AI summary

The invention encompasses a rapid and safe preparation method of sperm nuclei, improved sperm nuclei and a method of using the improved sperm nuclei to calibrate a flow cytometer with higher accuracy.