Cell Observation Optics Using Density Layers and Fluorescence
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Solution Overview
Problem
Existing cell observation methods struggle to accurately count cells in biological tissues containing fat due to difficulty in distinguishing between cells and lipid droplets, and centrifugation risks cell damage and loss.
Innovation Solution
An observation apparatus and method that separates a suspension into target and non-target layers based on specific gravity, uses non-coaxial observation light, and employs a filter to isolate and magnify cell images, excluding lipid droplets from the field of view.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If centrifugation is used to separate cells and fat, then cell counting accuracy is improved, but cell damage and cell loss occur
Solution Approach 1:
The suspension is segmented into multiple layers based on specific gravity differences, with the target layer containing cells separated from the non-target layer containing fat. This is achieved by allowing natural stratification in the holding unit without centrifugation, thus improving cell counting accuracy while avoiding cell damage
Solution Approach 2:
The fat component is extracted from the suspension by forming a separate non-target layer through specific gravity-based separation. This allows observation of cells in the target layer without interference from lipid droplets, achieving accurate cell counting without the need for centrifugation-induced cell damage
2Ease of operation
If bright field observation is used to count cells, then cell counting is simple, but cells and lipid droplets cannot be distinguished
Solution Approach 1:
Fluorescence labels are applied selectively to cells, giving them a distinct fluorescent property that differentiates them from non-fluorescent lipid droplets. This allows simple observation to become precise, as the fluorescent signal clearly identifies cells against the background of fat
Solution Approach 2:
The use of fluorescence introduces a new visual property (fluorescent emission) that distinguishes cells from lipid droplets. The fluorescent labels cause cells to emit light at specific wavelengths, creating a clear visual distinction that maintains operational simplicity while dramatically improving identification accuracy
3Measurement precision
If separation is performed to remove fat, then cell counting accuracy is improved, but cell loss occurs
Solution Approach 1:
A separation medium with specific gravity between that of cells and fat is introduced as an intermediary. This medium facilitates the formation of distinct layers where fat rises to the top and cells remain in the middle or lower layer, enabling accurate cell observation without direct contact or mechanical stress that would cause cell loss
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 cell counting without centrifugation, reducing cell damage and loss, and provides clearer images of cells through fluorescence observation.
Implementation Method 1
a holding unit that holds a suspension in a state of being separated into a target layer including target particles as an observation target among two or more kinds of particles included in the suspension and a non-target layer including particles other than the target particles among the two or more kinds of particles, the suspension including the two or more kinds of particles having different relative magnitudes of specific gravity with respect to a liquid
Implementation Method 2
an observation light source that causes observation light to be incident from a side of the target layer
Implementation Method 3
an objective lens that is provided on the side of the target layer and magnifies an image of the target particles
Implementation Method 4
The observation light source may output excitation light for exciting a fluorophore introduced into the target particles. The observation apparatus may have a filter that is provided between the objective lens and the imaging unit, blocks a wavelength component of the excitation light, and transmits a wavelength component of the fluorescence emitted from the target particles
Data Source
AI summary
Provided are an observation apparatus and an observation method capable of selectively observing target particles as an observation target among two or more kinds of particles included in a suspension.An observation apparatus includes: a holding unit that holds a suspension in a state of being separated into a target layer including target particles as an observation target among two or more kinds of particles included in the suspension and a non-target layer including particles other than the target particles among the two or more kinds of particles, the suspension including the two or more kinds of particles having different relative magnitudes of specific gravity with respect to a liquid; an observation light source that causes observation light to be incident from a side of the target layer; an objective lens that is provided on the side of the target layer and magnifies an image of the target particles; and an imaging unit that captures the image of the target particles magnified by the objective lens. A depth of field of an optical system including the objective lens is smaller than a thickness of the target layer.