Platelet Counting via Fluorescence Microscopy and Image Processing
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Solution Overview
Problem
Current methods for counting thrombocytes in blood samples, such as flow cytometry and centrifugation, require complex and expensive equipment, large blood volumes, and are not suitable for accurate counting in undiluted whole blood, often resulting in insufficient accuracy and the need for specialized laboratories.
Innovation Solution
A method involving specific fluorescent labeling of thrombocytes with antibodies and an agent inhibiting activation, followed by digital image processing under stationary fluorescence microscopy in a fluidic chamber, allowing for accurate counting without the need for extensive dilution or specialized equipment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If flow cytometry is used to count thrombocytes, then counting accuracy is improved, but equipment complexity and cost increase
Solution Approach 1:
The patent uses digital imaging to create a visual copy of the thrombocyte sample, allowing counting through image analysis rather than requiring complex flow cytometry equipment. The imaging system captures fluorescently labeled thrombocytes and computer algorithms automatically count them, replacing expensive specialized equipment with accessible imaging technology.
Solution Approach 2:
The patent replaces the mechanical and optical complexity of flow cytometry with a simpler imaging-based system. Instead of using flow cytometers with multiple lasers and detectors, the invention uses standard fluorescence microscopy combined with digital image processing to achieve accurate thrombocyte counting.
2Measurement precision
If flow cytometry is used to count thrombocytes, then counting accuracy is improved, but the volume of blood required increases
Solution Approach 1:
The patent segments the blood sample into a small volume loaded onto a microslide, which is then imaged. By dividing the counting task into discrete image fields rather than requiring analysis of large blood volumes through flow cytometry, the method achieves accurate counting with minimal blood sample (microliter scale).
3Measurement precision
If centrifugation is used to separate and count thrombocytes, then cell separation is improved, but device complexity and laboratory requirements increase
Solution Approach 1:
The patent replaces physical centrifugation separation with fluorescent labeling that visually distinguishes thrombocytes from other blood components. Instead of using centrifuges to separate cells by density, the invention uses fluorescence microscopy to selectively visualize and count thrombocytes based on their fluorescent labels, eliminating the need for complex separation equipment.
4Reliability
If thrombocytes are not inhibited from activation, then physiological state is maintained, but aggregation occurs reducing counting accuracy
Solution Approach 1:
The patent applies preliminary anti-action by adding activation inhibitors to the blood sample before imaging. These inhibitors prevent thrombocyte activation and aggregation that would otherwise occur during sample preparation, ensuring that thrombocytes remain in a countable state without requiring complex physiological maintenance systems.
Solution Approach 2:
The patent uses activation inhibitors as intermediary substances that mediate between the thrombocytes and the imaging process. These inhibitors temporarily modify thrombocyte behavior to prevent aggregation during counting, allowing accurate measurement without requiring the thrombocytes to be in a perfectly physiological state throughout the procedure.
5Ease of operation
If high dilution is applied for flow cytometry, then cell passage is improved, but the volume of blood required increases
Solution Approach 1:
The patent transitions from the flow dimension of flow cytometry to a static imaging dimension. Instead of requiring diluted cell suspensions to pass through a flow chamber one by one, the invention loads a small volume of undiluted or minimally diluted blood onto a microslide and images the thrombocytes in place, eliminating the need for high dilution factors while maintaining ease of operation.
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 reliable and efficient counting of thrombocytes in small blood volumes with high accuracy, reducing the need for large equipment and specialized settings, while preventing activation and aggregation of platelets, thus providing a simple and cost-effective solution for platelet quantification.
Implementation Method 1
fluorescent markers capable of binding specifically with thrombocytes; the acquisition of at least one digital image (IN) of said mixture by fluorescence microscopy under stationary lighting
Data Source
Figure 1~2
AI summary
A method for counting platelets contained in a blood sample, comprising: - mixing said sample with: ○ fluorescent markers capable of binding specifically to platelets; and ○ an agent inhibiting the activation of said platelets; - introducing the sample (E) into a fluidic chamber (CF) having at least one transparent face; - acquiring at least one digital image (IN) of said sample by fluorescence microscopy under stationary illumination; and - counting the platelets (T) present in said image or in each image by computer image processing means (O). Apparatus for carrying out this method.