Reticulocyte and Platelet Simulating Particles With Stable Fluorescent RBCs
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for preparing reticulocyte and platelet simulating particles are complex, costly, and inefficient, failing to accurately simulate human blood cell characteristics, leading to inaccurate hematology analyzer results.
Innovation Solution
A method involving staining mammalian anucleated red blood cells with a protein fluorescent dye activated by N-hydroxysuccinimide, followed by fixing, to create reticulocyte and platelet simulating particles with volumes of 60-120 fL and 2-25 fL respectively, ensuring similar membrane properties and fluorescence characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If human platelets are used as raw materials with polyethylene glycol to enhance stability, then stability is improved, but platelets are easily activated causing aggregation effect and high cost
Solution Approach 1:
The patent replaces expensive human platelets with inexpensive animal red blood cells as the base material for simulating particles. This substitution eliminates the aggregation problem inherent in human platelets while maintaining the ability to simulate platelet characteristics through fluorescent staining and controlled deformation, thereby resolving the contradiction between stability and aggregation effect.
Solution Approach 2:
The patent creates artificial copies of platelet simulating particles using animal red blood cells that are stained with fluorescent dyes and subjected to controlled osmotic pressure changes. These copies replicate the optical and morphological characteristics of human platelets without possessing the biological limitations of actual platelets, thus achieving stability without aggregation.
2Manufacturing precision
If goat red blood cells are shrunk to simulate human platelet volume distribution, then volume simulation is improved, but cells have tendency to recover from deformation in isosmotic preservation solutions
Solution Approach 1:
The patent employs controlled osmotic pressure changes during the preparation process to deform animal red blood cells into platelet-like shapes and sizes. By carefully adjusting the osmotic parameters and using non-isosmotic preservation solutions, the deformed cells maintain their simulated platelet morphology without recovering, thus achieving both volume precision and stability.
3Manufacturing precision
If multiple goat blood types are mixed to simulate human platelet volume distribution, then volume distribution is improved, but process complexity increases
Solution Approach 1:
Instead of mixing multiple blood sources, the patent achieves diverse platelet volume distributions by applying controlled osmotic pressure treatments to a single animal red blood cell source. This parameter-based approach allows precise control over cell deformation and final volume distribution, simplifying the process while maintaining manufacturing precision.
4Manufacturing precision
If red blood cells are swollen by changing osmolarity to permit nucleic acid entry, then reticulocyte morphology simulation is improved, but cell membrane strength decreases and mass production efficiency is low
Solution Approach 1:
Instead of swelling red blood cells to create reticulocyte-like structures, the patent uses normally sized animal red blood cells and deforms them through controlled osmotic pressure to create reticulocyte simulating particles. This inverted approach maintains cell membrane integrity while achieving the desired morphology, enabling high-efficiency mass production.
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 method produces stable simulating particles that accurately mimic reticulocytes and platelets, maintaining cell activity and stability, and do not interfere with other blood cell counts, facilitating efficient and accurate hematology reference controls.
Implementation Method 1
staining mammalian anucleated red blood cells with a protein fluorescent dye having a carboxyl group activated by N-hydroxysuccinimide
Implementation Method 2
followed by fixing, to create reticulocyte and platelet simulating particles
Data Source
AI summary
Provided are a method for preparing reticulocyte simulating particles and platelet simulating particles, and a reference control. The method for preparing the reticulocyte simulating particles comprises: staining mammalian anucleated red blood cells having a volume of 60-120 fL with a protein fluorescent dye activated by N-hydroxysuccinimide, and fixing the anucleated red blood cells to prepare the reticulocyte simulating particles. The platelet simulating particles are prepared from mammalian anucleated red blood cells having a volume of 2-25 fL, and the steps of preparing the platelet simulating particles are the same as that for the reticulocyte simulating particles. The preparation method comprises: using an protein fluorescent dye activated by N-hydroxysuccinimide to stain mammalian anucleated red blood cells having different volumes, so as to respectively obtain reticulocyte simulating particles and platelet simulating particles.