Formaldehyde-Free Parasite Fixative Composition for Reliable Staining

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Solution Overview

Problem

Existing fixative solutions for biological samples, particularly parasitological samples, are hazardous and struggle to effectively kill organisms like helminths with tough outer layers, and their impact on staining properties is unpredictable.

Innovation Solution

A fixative solution comprising C2 to C6 carboxylic acid, C2 to C6 carboxylate salt or halide salt, and optionally biocidal essential oils, which rapidly kills organisms and preserves cellular structure without toxic additives, facilitating good staining properties.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If formaldehyde is used in fixative solutions, then fixation effectiveness is improved, but toxicity and carcinogenicity increase

Engineering Contradiction:
Improvefixation effectivenessVSAvoidtoxicity
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the chemical parameters of the fixative solution by replacing formaldehyde with a specific combination of carboxylic acid (acetic acid at 0.5-3.5M), carboxylate salt, and halide salt (calcium chloride/magnesium chloride at 0.02-0.5M). This parameter substitution maintains fixation effectiveness while eliminating the toxic and carcinogenic properties of formaldehyde.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite fixative system combining multiple components: carboxylic acid, carboxylate salt, and halide salt. This composite formulation works synergistically to achieve effective fixation of parasitic organisms without requiring toxic substances like formaldehyde, thereby maintaining reliability while reducing harmful factors.

Inventive Principle:
Principle #40Composite materials

2Reliability

If highly toxic chemicals are used to kill parasitical organisms, then killing effectiveness is improved, but safety hazards increase

Engineering Contradiction:
Improvekilling effectivenessVSAvoidsafety hazards
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the chemical composition parameters by using non-hazardous carboxylic acid and halide salt combinations at specific concentrations. This allows achieving effective killing of parasitic organisms with tough outer layers without the safety hazards associated with highly toxic chemicals, while maintaining reliable fixation.

Inventive Principle:
Principle #35Parameter changes

3Object-affected harmful factors

If non-hazardous fixatives are used, then safety is improved, but killing effectiveness against resilient organisms decreases

Engineering Contradiction:
ImprovesafetyVSAvoidkilling effectiveness
Core Design Contradiction:
Object-affected harmful factorsVSReliability

Solution Approach 1:

The patent develops a composite non-hazardous fixative system combining carboxylic acid, carboxylate salt, and halide salt that works together to effectively kill resilient parasitic organisms. This composite formulation achieves reliable killing effectiveness while maintaining safety by avoiding toxic additives, thus resolving the contradiction between safety and killing effectiveness.

Inventive Principle:
Principle #40Composite materials

4Manufacturing precision

If fixative formulation is modified, then staining properties may be improved, but predictability of staining results decreases

Engineering Contradiction:
Improvestaining propertiesVSAvoidpredictability
Core Design Contradiction:
Manufacturing precisionVSLoss of information

Solution Approach 1:

The patent establishes specific parameter ranges for the fixative components (carboxylic acid at 0.5-3.5M, carboxylate salt at 0.02-0.5M, halide salt at 0.02-0.5M) that have been optimized to produce consistent and predictable staining results. By defining precise concentration ranges rather than allowing arbitrary modifications, the patent maintains both improved staining properties and result predictability.

Inventive Principle:
Principle #35Parameter changes

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

The solution achieves rapid organism killing, preserves cellular structure, and ensures excellent staining properties, even after prolonged periods, making it suitable for microscopic examination.

Implementation Method 1

a C2 to C6 carboxylic acid at a concentration of 0.5M or greater... It has been found that such concentrations of carboxylic acid (particularly acetic acid) result in a rapid killing of biological organisms such as helminth ova

Methodology Applied
Scientific EffectCellular disruption:

Implementation Method 2

The carboxylate salt or halide salt of component b) may for example be the salt of an alkali or alkaline earth metal or of ammonium other cation that confers reasonable solubility... It has been found that such halide salts help preserve the cellular structure, possibly an osmoregulatory effect involving the halide salts counteracting the turgor effect of the carboxylic acid

Methodology Applied
Scientific EffectOsmoregulatory effect: Osmotic Pressure

Data Source

PatentUS12601660B2Fixative solution and method of preparation of biological sample
Publication Date: 2026.04.14 APACOR

AI summary

A fixative contains carboxylic acid, preferably acetic acid, for parasitological samples. An exemplary aqueous fixative solution for parasitological samples comprises: a) a C2 to C6 carboxylic acid (preferably acetic acid) at a concentration of 0.5M or greater; and b) a C2 to C6 carboxylate salt or halide salt at a concentration of 0.02M or greater, preferably 0.04M or greater. An optional composition comprises 1.3M acetic acid, 12 mM sodium benzoate, 0.9% calcium chloride, 0.9% calcium bromide, 100 μg/ml menthol and 350 μg/ml thymol. The fixative is free of formaldehyde or other hazardous compounds, and exhibits good killing and staining properties when used with parasitological samples such as ova and helminths.