Single-Step Blood Cell Staining Composition for Rapid Differentiation
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Solution Overview
Problem
Current blood cell staining techniques require multiple steps and significant time, leading to inefficiencies in automated analysis systems, and struggle with even staining and differentiation of cell types like immature granulocytes.
Innovation Solution
A particle contrast agent composition comprising Basic Fuchsin, Brilliant Cresyl Blue, Azure B, Crystal Violet, and New Methylene Blue, combined with permeabilizing and fixing agents, is used to stain blood samples in a single-step process, suitable for automated flow cytometry systems.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If traditional multi-step staining process is used, then cell structure preservation and staining quality are improved, but processing time and system complexity increase
Solution Approach 1:
The patent combines multiple staining agents (Wright's stain components including methylene blue, eosin, and other dyes) into a single composite staining solution. This merging of multiple staining steps into one simultaneous application reduces processing time while maintaining the differential staining quality needed to distinguish various blood cell types and their granules
Solution Approach 2:
The single staining composition serves multiple functions simultaneously: it stains different cell types (neutrophils, eosinophils, basophils, lymphocytes, monocytes), differentiates granule types (specific and nonspecific granules), and preserves cell morphology all in one step, replacing the need for multiple sequential staining operations
2Manufacturing precision
If traditional multi-step staining process is used, then staining quality is improved, but device complexity and operation difficulty increase
Solution Approach 1:
The patent merges multiple separate staining solutions and application steps into a single composite staining composition and single application step. This eliminates the complexity of managing multiple reagents, timing sequences, and transfer operations while maintaining comprehensive staining capability for all blood cell types
Solution Approach 2:
The universal staining composition performs all necessary staining functions in one application, making the process easier to operate and reducing the complexity of the staining system while achieving differential staining of all major blood cell types and their granules
3Productivity
If rapid staining is performed, then processing speed is improved, but staining evenness and cell differentiation capability deteriorate
Solution Approach 1:
The patent optimizes the chemical parameters of the staining composition, including pH, ionic strength, and dye concentrations, to enable rapid penetration and binding. The composition is formulated with specific ratios of acidic and basic dyes that allow quick differential staining while maintaining even color distribution across all cell types within seconds
4Productivity
If sample volume is reduced for automated systems, then system efficiency is improved, but staining completeness and cell differentiation capability worsen
Solution Approach 1:
The staining composition is formulated with enhanced solubility and stability parameters that allow effective staining in reduced sample volumes. The optimized dye concentrations and buffer composition ensure complete cell penetration and granule differentiation even when working with the small sample volumes typical of automated hematology analyzers
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 composition achieves rapid and even staining of blood cells, allowing for efficient differentiation and sub-categorization of cell types within 90 seconds, enhancing the speed and accuracy of automated analysis.
Implementation Method 1
The particle contrast agent composition can include: a first staining agent, Basic Fuchsin; a second staining agent, at least one of Brilliant Cresyl Blue, Azure B, Crystal Violet and New Methylene Blue
Implementation Method 2
a permeabilizing agent in an amount effective to permeabilize the cell membrane of the white blood cells
Implementation Method 3
a fixing agent in an amount effective to fix the white blood cells
Data Source
AI summary
The present disclosure relates to a staining methodology employing a particle contrast agent composition. The compositions described herein are capable of rapidly staining cells in a single step. The particle contrast agent composition can be comprised of a combination of one or more particle contrast agents, one or more permeabilizing agents, and one or more fixing agents. The particle contrast agent composition can include Basic Fuchsin; at least one of Brilliant Cresyl Blue, Azure B, Crystal Violet and New Methylene Blue; one or more permeabilizing agents; and Gluteraldehyde.