Fluid Layering Device for Density Gradient Overlay
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Solution Overview
Problem
The existing density gradient protocol for isolating leukocytes (white blood cells) is time-consuming, messy, and requires extensive hands-on training, particularly due to the challenging process of overlaying blood or cellular suspension over a density gradient in centrifuge tubes.
Innovation Solution
A fluid layering device that includes a reservoir for storing the overlay fluid, a fluid barrier separating the reservoir and the container, fluid channels across the fluid barrier, and an exhaust regulator, allowing for a controlled, steady flow of blood or cellular suspension onto the density gradient without disturbing its surface.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the overlay method is performed manually by pouring blood or cellular suspension over the density gradient, then the isolation of leukocytes can be achieved, but the process is time-consuming and requires extensive hands-on training
Solution Approach 1:
A fluid barrier is introduced as an intermediary component between the reservoir containing blood or cellular suspension and the centrifuge tube containing the density gradient. This barrier controls the flow of fluids to enable automated overlay without manual pouring, reducing time and training requirements while maintaining isolation yield
Solution Approach 2:
The device enables self-service operation where the system automatically performs the overlay process without requiring skilled manual intervention. The automated fluid control mechanisms handle the entire overlay sequence, eliminating the need for extensive hands-on training
2Reliability
If manual overlay is performed by pouring fluids, then leukocyte isolation can be achieved, but spills and mixing of fluids frequently occur
Solution Approach 1:
The fluid barrier acts as a mediator that precisely controls fluid discharge, preventing spills and mixing that occur during manual pouring. It regulates the overlay process to maintain distinct fluid layers without requiring high operator skill
Solution Approach 2:
The manual mechanical pouring action is replaced with a controlled automated system using the fluid barrier and regulated flow mechanisms. This substitution eliminates the variability and errors associated with manual technique while maintaining precise fluid layering
3Productivity
If the overlay process is performed manually, then leukocyte isolation can be completed, but the process is tedious and limits the volume of samples that can be processed
Solution Approach 1:
The automated device performs the overlay process without requiring continuous hands-on intervention. Multiple samples can be processed sequentially or in parallel, significantly increasing productivity while reducing the duration of hands-on time required per sample
Solution Approach 2:
The device allows preliminary preparation of samples and automated execution of the overlay process. This enables high-throughput processing where multiple samples can be loaded and processed with minimal ongoing manual intervention, increasing overall productivity
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 device eliminates the need for prior training, reduces human error, and significantly speeds up the overlay process, while ensuring a perfect and uniform layering of fluids, thereby increasing the yield and allowing for larger experiments to be performed efficiently.
Implementation Method 1
a fluid barrier configured to prevent passage of fluid from a reservoir toward a bottom portion of the centrifuge tube
Implementation Method 2
a gas vent configured to vent from the centrifuge tube gas displaced by the controlled flow of overlay fluid
Implementation Method 3
a volume of a base material, such as Ficoll or any other suitable density gradient
Implementation Method 4
The isolation and preparation of leukocytes, more generally referred to as 'white blood cells' (WBC's), from whole blood or cellular suspension using a density gradient
Data Source
AI summary
A device is described that overlays a first fluid, such as blood or a cellular suspension onto a base material, such as a density gradient. In some embodiments, the fluid layering device includes a cylindrical reservoir, a fluid barrier, a coupling extension, a plunger, and an exhaust vent. The fluid layering device can be coupled through its coupling extension to an open end of a container, such as a conical centrifuge tube, including the density gradient. Once attached, the plunger may be lowered to a position above the surface of the density gradient. A first fluid may flow from the reservoir into the conical tube across the plunger, so that a suitable overlay is formed without substantially disturbing a surface of the density gradient.