研究目的
To understand complex and dynamic biological processes by analyzing cellular responses with single-cell sensitivity using noninvasive optical methods.
研究成果
The label-free optical approach can distinguish between close cell types and fine cellular states, providing high classification sensitivities. The method is robust to uncontrollable factors and can identify individual mice that produce cell populations with outlier variances.
研究不足
The study is limited by the complexity of disentangling the effect of different parameters on the data variability and the potential influence of instrument drifts on spectral variables.
1：Experimental Design and Method Selection:
A noninvasive optical approach combining quantitative phase imaging and Raman spectroscopy measurements was used to retrieve quantitative biomarkers of both morphological and molecular phenotypes of individual cells.
2：Sample Selection and Data Sources:
Primary cultured peritoneal cavity macrophages and a macrophage cell line (Raw264) were used, exposed to lipopolysaccharide (LPS) to manifest phenotypic changes.
3：List of Experimental Equipment and Materials:
A multimodal microscope with a 40x microscope objective, a Mach-Zehnder interferometer illuminated with a laser diode, and a spectrometer to measure the vibrational spectrum.
4：Experimental Procedures and Operational Workflow:
Cells were imaged with quantitative phase imaging and Raman spectroscopy simultaneously, and auto-fluorescence images were acquired in a standard epi-fluorescence configuration.
5：Data Analysis Methods:
Data was analyzed using principal component analysis (PCA) and penalised logistic regression for classification.
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multimodal microscope
Imaging and spectroscopy of cells
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40x microscope objective
Magnification and imaging of cells
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Mach-Zehnder interferometer
Quantitative phase imaging
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laser diode
Illumination for imaging
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spectrometer
Measurement of vibrational spectrum
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scientific CMOS detector
Detection of Raman spectra
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mercury lamp
Excitation for auto-fluorescence imaging
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DAPI filter set
Filtering for auto-fluorescence imaging
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