Abstract
Qualifications of intracellular structure were performed for the first time using the gray-level co-occurrence matrix (GLCM) method for images of cells obtained by resolution-enhanced photothermal imaging. The GLCM method has been used to extract five parameters of texture features for five different types of cells in mouse brain; pyramidal neurons and glial cells in the basal nucleus (BGl), dentate gyrus granule cells, cerebellar Purkinje cells, and cerebellar granule cells. The parameters are correlation, contrast, angular second moment (ASM), inverse difference moment (IDM), and entropy for the images of cells of interest in a mouse brain. The parameters vary depending on the pixel distance taken in the analysis method. Based on the obtained results, we identified that the most suitable GLCM parameter is IDM for pyramidal neurons and BGI, granule cells in the dentate gyrus, Purkinje cells and granule cells in the cerebellum. It was also found that the ASM is the most appropriate for neurons in the basal nucleus.
Highlights
Optical microscopy has been extensively used for biological imaging for many years but the resolution is limited by diffraction.[1]
We performed for the first time qualifications of intracellular structure in the resolution-enhanced images of cells in mouse brain obtained by PT imaging, which we developed to perform nondiffraction limited imaging using a compact system without requiring fluorescent capability.[16,17,18]
The experimental resolution is defined as the FWHM of Gaussian function obtained by the least squares mean (LSM) fitting to the observed intensity distribution of the gold nanoparticles (GNPs), and the values following “Æ” symbol are the standard deviations obtained by the LSM
Summary
Optical microscopy has been extensively used for biological imaging for many years but the resolution is limited by diffraction.[1] Recently, in order to visualize a variety of nanoscale cellular components with high specificity and high spatial resolution, several superresolution imaging techniques such as stimulated emission depletion microscopy, structured illumination microscopy, and stochastic optical reconstruction microscopy have been developed reaching sub-100-nm resolution.[2,3,4,5,6,7] Such high resolution has revealed fine intracellular structures Another direction of the development of microscope is the imaging of nonfluorescent system with high sensitivity and resolution using photothermal (PT) effects with resolution enhancement.[8,9,10,11,12] In these studies, the high resolution capability thanks to the pump–probe scheme has revealed fine intracellular structures, intracellular “global” statistical features of the intracellular structure such as uniformity, correlation among the microstructures, and entropic randomness have not been investigated. The accurate detection of a deformation or structural changes at the cellular level depends on the efficiency of the microscope and the skill of the
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