Abstract
The visualization of heterogeneous morphology, segmentation and quantification of image features is a crucial point for nonlinear optics microscopy applications, spanning from imaging of living cells or tissues to biomedical diagnostic. In this paper, a methodology combining stimulated Raman scattering microscopy and image analysis technique is presented. The basic idea is to join the potential of vibrational contrast of stimulated Raman scattering and the strength of imaging analysis technique in order to delineate subcellular morphology with chemical specificity. Validation tests on label free imaging of polystyrene-beads and of adipocyte cells are reported and discussed.
Highlights
A subject of wide interest in the physical and life sciences is the noninvasive characterization of microscopic objects within a complex heterogeneous system through nonlinear optical microscopy [1]
The intensity of the pump beam is modulated with an electro-optic modulator and the modulation transfer to the probe beam is measured with a lock-in amplifier (LIA), after blocking the pump beam with an optical filter
In order to validate the proposed methodology, two examples are reported and discussed. In the former, label free imaging of polystyrene beads is considered, while in the latter, label free imaging of fixed adipocyte cells is carried out
Summary
A subject of wide interest in the physical and life sciences is the noninvasive characterization of microscopic objects within a complex heterogeneous system through nonlinear optical microscopy [1]. The basic idea is that images of cells tend to give only an intuitive understanding of the structure and the spatial distributions of chemicals and organelles, while identification and quantification of such parameters are necessary in order to accurately compare images and make objective conclusions about an experiment. This is crucial for a number of applications spanning from imaging of living cells or tissues until biomedical diagnostic [2]
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