Infrared Spectroscopy for Cell Viability Detection
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Solution Overview
Problem
Conventional methods for determining whether organic matter is present and whether organisms are alive face challenges such as the need for costly reagents, labor-intensive processes, difficulty in quantification, and inability to monitor changes over time, often resulting in errors and inefficiencies.
Innovation Solution
An apparatus and method utilizing infrared absorption spectroscopy to detect amide infrared absorption peaks, specifically the amide I, amide II, and amide III peaks, to determine the presence and viability of organisms by analyzing changes in wave number values and peak shifts, eliminating the need for reagents and enabling continuous monitoring of the same sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If MTT analysis method is used to measure apoptosis, then reproducible optical density can be obtained with minimum physical treatment, but costly reagents are required and a lot of measurement time and labor are required
Solution Approach 1:
The patent replaces the chemical-based MTT assay system with an infrared spectroscopic system. Instead of using MTT reagents that require chemical reactions and subsequent measurement, the invention uses infrared light to directly detect amide bond vibrations in proteins, providing reproducible measurements without chemical reagents and significantly reducing measurement time and labor
Solution Approach 2:
The infrared spectroscopic method allows samples to provide their own measurement signal through the natural infrared absorption characteristics of amide bonds in proteins. No external reagents or labels are needed - the sample's inherent molecular vibrations serve as the measurement signal, eliminating the need for costly reagents and complex preparation procedures
2Ease of operation
If MTT analysis method is used, then optical density measurement is simple, but extracellular abiotic factors may cause fatal errors in analysis results
Solution Approach 1:
The patent replaces the chemical reduction-based MTT assay with a physical infrared spectroscopic method. Instead of measuring chemical reaction products that can be influenced by abiotic factors, the invention directly detects the vibrational states of amide bonds in proteins, providing reliable measurements that are not susceptible to interference from extracellular abiotic factors
Solution Approach 2:
The infrared spectroscopic method uses amide bond vibrations as an intermediary signal that directly reflects protein content and cellular state. This intermediary provides a more reliable measurement pathway compared to MTT's chemical reaction pathway, as it bypasses the vulnerable step of chemical reduction that can be affected by abiotic factors
3Loss of information
If flow cytometry is used for cell analysis, then detailed cell information can be obtained, but irreversible pretreatment is required involving sample labeling and cell detachment
Solution Approach 1:
The infrared spectroscopic method allows adherent cells to be measured in their native state without detachment or labeling. The cells' own infrared absorption signals provide sufficient information for viability assessment, eliminating the need for complex pretreatment while maintaining measurement capability
Solution Approach 2:
The patent replaces the mechanical and chemical processes of flow cytometry (detachment, labeling, suspension) with a non-invasive infrared spectroscopic measurement. This substitution maintains the ability to obtain detailed cell information while completely eliminating the irreversible pretreatment steps
4Measurement precision
If specific protein detection method is used to measure apoptosis, then protein expression amount can be quantified, but many samples and experimental procedures must be prepared and performed
Solution Approach 1:
The infrared spectroscopic method provides a universal measurement approach that can assess cell viability and protein content through a single measurement process. Instead of requiring separate procedures for different protein detections, the infrared spectrum's amide bond region provides comprehensive information about protein content and cellular state in one experiment
Solution Approach 2:
The patent merges multiple measurement objectives (cell viability, protein content, cellular state) into a single infrared spectroscopic measurement. The amide bond absorption signals in the infrared spectrum simultaneously provide information about protein concentration and conformational changes, eliminating the need for multiple separate experimental procedures
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
This approach allows for simple, cost-effective, and accurate measurement of organic matter and organism viability without reagents, overcoming previous method limitations by enabling quantification and continuous monitoring of cell changes, thereby reducing errors and labor costs.
Implementation Method 1
irradiating a sample with infrared rays and detecting the infrared rays transmitted through or reflected from the sample
Implementation Method 2
characteristic whereby an infrared absorption spectrum changes depending on whether organic matter is present or whether or not organisms are alive
Implementation Method 3
identifying an amide infrared absorption peak of the sample using the detected infrared rays
Data Source
AI summary
This invention relates to an apparatus and a method for measuring whether organic matter is present or whether or not organisms (cells or tissue) are alive using infrared absorption spectroscopic analysis. The measurement apparatus of the invention includes an infrared light source for radiating infrared rays on a sample, an infrared detection unit for detecting the infrared rays transmitted or reflected from the sample, and a determination unit for identifying an amide infrared absorption peak of the sample using the detected infrared rays and for determining whether organic matter is present or whether organisms are alive or not in the sample using the identified amide infrared absorption peak. In this invention, a reagent is not used, simple measurement is performed, quantification is feasible, and the presence or absence of cells or tissues and changes in the life and death can be consecutively measured for the same sample.