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Scattering Inversion Study for Suspended Label-Free Lymphocytes with Complex Fine Structures
Lu Zhang1, Huijun Wang1, Jianyi Liu2
1Xi'an Jiaotong University, School of Mechanical Engineering, State Key Laboratory for Manufacturing Systems Engineering, Xi'an 710049, China.
Insights
This study introduces an inverse light scattering (ILS) method to measure lymphocyte volumes, aiding in pathological diagnosis. The technique accurately distinguishes malignant from normal cells without damaging samples.
Area of Science:
- Biophysics
- Pathology
- Optical methods
Background:
- Light scattering provides cell morphology information linked to biophysical states.
- Understanding the relationship between complex cell structures and scattering characteristics is crucial for pathological analysis.
Purpose of the Study:
- To develop and validate an inverse light scattering (ILS) method for label-free, quantitative analysis of suspended lymphocyte volumes.
- To identify pathological states by distinguishing malignant from normal lymphocytes based on their morphology.
Main Methods:
- Modeled clinical lymphocyte morphologies using Gaussian random sphere geometry and confocal scanning data.
- Developed an ILS approach to quantitatively inverse the volumes of cell membranes and nuclei.
- Investigated the specificity of ILS for complex structures, including surface roughness, posture, and refractive index.
Main Results:
- Demonstrated a linear relationship between membrane/nucleus volumes and forward scattering image properties (effective area and entropy).
- Achieved specificity deviations below 3.5% for the ILS method.
- Validated the method with microspheres and clinical leukocytes, showing high Pearson product-moment correlation coefficients (PPMCC) up to 0.9926.
Conclusions:
- The proposed ILS method effectively identifies suspended, label-free lymphocytes.
- This technique offers a non-destructive approach for pathological examination without complex experimental setups.
Abstract:
Objective and Impact Statement. Distinguishing malignant lymphocytes from normal ones is vital in pathological examination. We proposed an inverse light scattering (ILS) method for label-free suspended lymphocytes with complex fine structures to identify their volumes for pathological state. Introduction. Light scattering as cell's "fingerprint" provides valuable morphology information closely related to its biophysical states. However, the detail relationships between the morphology with complex fine structures and its scattering characters are not fully understood. Methods. To quantitatively inverse the volumes of membrane and nucleus as the main scatterers, clinical lymphocyte morphologies were modeled combining the Gaussian random sphere geometry algorithm by 750 reconstructed results after confocal scanning, which allowed the accurate simulation to solve ILS problem. For complex fine structures, the specificity for ILS study was firstly discussed (to our knowledge) considering the differences of not only surface roughness, posture, but also the ratio of nucleus to the cytoplasm and refractive index. Results. The volumes of membrane and nucleus were proved theoretically to have good linear relationship with the effective area and entropy of forward scattering images. Their specificity deviations were less than 3.5%. Then, our experimental results for microsphere and clinical leukocytes showed the Pearson product-moment correlation coefficients (PPMCC) of this linear relationship were up to 0.9830~0.9926. Conclusion. Our scattering inversion method could be effectively applied to identify suspended label-free lymphocytes without destructive sample pretreatments and complex experimental systems.
