Reticulocyte Identification Using Transparent Panel Optical Density
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Solution Overview
Problem
Existing blood analysis methods require precise sample volumes and often involve dilution or staining, leading to inaccuracies and are not suitable for determining red blood cell indices in species like birds, reptiles, and fish, where red blood cells are nucleated or small, making it difficult to obtain accurate complete blood counts.
Innovation Solution
A method using an analysis chamber where a substantially undiluted blood sample is imaged with transparent panels, allowing red blood cells to contact both surfaces, enabling optical density determination for calculating cell volume and hemoglobin concentration without external dyes or diluents, suitable for small sample volumes and adaptable for handheld devices.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If manual or centrifugal methods are used for blood cell analysis, then accurate cell enumeration can be achieved, but precise sample volume dispensing is required which increases device complexity and is not suitable for point-of-care settings
Solution Approach 1:
The patent replaces mechanical impedance-based counting with optical imaging and analysis. Instead of using impedance methods that require precise sample volume dispensing and complex mechanical systems, the invention uses transparent panel imaging to directly visualize and count cells, eliminating the need for precise volumetric control while maintaining counting accuracy
Solution Approach 2:
The patent creates an optical copy or image of the blood sample between transparent panels. By capturing an image of the cells sandwiched between the panels and analyzing this optical replica, the system eliminates the need for direct manipulation and precise measurement of the actual sample volume, simplifying the device while maintaining measurement accuracy
2Extent of automation
If impedance or flow cytometric methods are used, then automated cell counting is achieved, but the methods require dilution with reagents and calibration which increases device complexity and waste generation
Solution Approach 1:
The patent enables the blood sample to serve itself by sandwiching it between transparent panels for direct imaging. The cells naturally position themselves between the panels without requiring external dilution reagents or complex preparation steps. The transparent panels themselves serve as both the containment structure and the imaging medium, eliminating the need for separate calibration standards or dilution protocols
Solution Approach 2:
The patent extracts the essential function of cell visualization from complex fluid handling systems. By removing the need for dilution reagents, calibration standards, and complex flow systems, the invention isolates the core function of cell counting to a simple transparent panel sandwich structure that can be imaged and analyzed directly
3Ease of operation
If peripheral smear methods are used, then no precise volume dispensing is needed, but a high degree of training is required for proper examination which reduces ease of operation
Solution Approach 1:
The patent replaces the subjective visual interpretation required in peripheral smear analysis with objective optical imaging and automated analysis. Instead of relying on trained personnel to visually examine and interpret smears, the system captures digital images and uses automated algorithms to identify and count cells, eliminating the need for extensive training while maintaining ease of operation
Solution Approach 2:
The patent creates a digital optical copy of the blood sample between transparent panels that can be analyzed objectively. This digital replica allows for automated image processing and cell identification algorithms to replace human visual inspection, eliminating the need for trained operators while preserving the simplicity of direct sample placement
4Quantity of substance
If centrifugal separation methods are used, then sample separation into layers is achieved, but large volumes of contaminated waste are generated which increases loss of substance
Solution Approach 1:
The patent extracts only the essential function of cell visualization without requiring sample separation into layers. By using transparent panels to directly image whole blood or diluted samples, the system eliminates the centrifugation step that generates large volumes of waste, while still enabling cell identification and counting through optical means
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
Enables accurate determination of red blood cell indices, including volume and hemoglobin content, in small sample volumes directly from patients, without the need for external reagents or calibration, and is operable in various orientations and environments, including microgravity.
Implementation Method 1
an analysis chamber adapted to quiescently hold the sample for analysis, the chamber defined by an interior surface of a first panel, and an interior surface of a second panel, wherein both panels are transparent
Implementation Method 2
imaging the at least one red blood cell contacting the interior surfaces; determining an optical density of a portion of the imaged red blood cell
Data Source
AI summary
A method and apparatus for identifying reticulocytes within a blood sample is provided. The method includes the steps of: a) depositing the sample into an analysis chamber adapted to quiescently hold the sample for analysis, and the chamber has a known or determinable height extending between the interior surfaces of panels, which height is such that at least one red blood cell, or an aggregate of red blood cells, within the sample contacts both of the interior surfaces; b) admixing a supravital dye with the sample, which dye is operable to cause reticulin to fluoresce when excited by light of one or more predetermined wavelengths; c) imaging the sample using light that includes the one or more predetermined wavelengths that cause reticulin to fluoresce; d) imaging the sample using light that is absorbed by hemoglobin to produce values of optical density on a per image unit basis; and e) identifying reticulocytes within the sample using the image of the sample created with light that causes the dyed reticulin to fluoresce, and using the per image unit optical density values.