Multimodality of an optical system implies the use of one or more optical techniques to improve the system's overall performance and maximum utility. In this article, we demonstrate a multimodal system with oblique illumination that combines two different techniques; fluorescence micro‐endoscopy and spectroscopy simultaneously and can be utilized to obtain diverse information from the same location of biological sample. In present system, use of graded index (GRIN) rod‐lens makes it highly compact and oblique incidence decouples illumination geometry with collection geometry, preventing CCD cameras from saturation and reduces number of optical elements, thereby making system further miniaturized and field‐portable. It also overcomes the disadvantages of undesired reflections from different optical elements. The experimental results of simultaneous imaging and spectroscopy of the biological samples are presented along with quantitative spectroscopic parameters; peak wavelength shift, area under the curve and full width half maximum (FWHM). The spatial resolution, spectral resolution and field of view of the system are found to be 4.38 μm, 0.5 nm and 2.071×1.5480.25emmm2, respectively. Furthermore, we have obtained the red shift for cancerous oral tissue with respect to normal oral tissue 5.79 ± 1.071 nm. This could be important indicator for oral cancer screening.
In this paper, we demonstrate the white light phase shifting interferometer employed as whole slide scanner and phase profiler for determining qualitative and quantitative information over large field-of-view (FOV). Experiments were performed on human erythrocytes and MG63 Osteosarcoma cells. Here, we have recorded microscopic images and phase shifted white light interferograms simultaneously in a stepped manner. Sample slide is translated in transverse direction such that there exists a correlation between the adjacent frames, and they were stitched together using correlation functions. Final stitched image has a FOV of 0.24 Â 1.14 mm with high resolution ~0.8 μm. Circular Hough transform algorithm is implemented to the resulting image for cell counting and five-step phase shifting algorithm is utilised to retrieve the phase profiles over a large FOV. Further, this technique is utilised to study the difference between normal and anaemic erythrocytes. Significant changes are observed in anaemic cells as compared to normal cells.
Objective
Fluorescence‐based methods are highly specific and sensitive and have potential in breast cancer detection. Simultaneous fluorescence imaging and spectroscopy during intraoperative procedures of breast cancer have great advantages in detection of tumor margin as well as in classification of tumor to healthy tissues. Intra‐operative real‐time confirmation of breast cancer tumor margin is the aim of surgeons, and therefore, there is an urgent need for such techniques and devices which fulfill the surgeon's priorities.
Methods
In this article, we propose the development of fluorescence‐based smartphone imaging and spectroscopic point‐of‐care multi‐modal devices for detection of invasive ductal carcinoma in tumor margin during removal of tumor. These multimodal devices are portable, cost‐effective, noninvasive, and user‐friendly. Molecular level sensitivity of fluorescence process shows different behavior in normal, cancerous and marginal tissues. We observed significant spectral changes, such as, red‐shift, full‐width half maximum (FWHM), and increased intensity as we go towards tumor center from normal tissue. High contrast in fluorescence images and spectra are also recorded for cancer tissues compared to healthy tissues. Preliminary results for the initial trial of the devices are reported in this article.
Results
A total 44 spectra from 11 patients of invasive ductal carcinoma (11 spectra for invasive ductal carcinoma and rest are normal and negative margins) are used. Principle component analysis is used for the classification of invasive ductal carcinoma with an accuracy of 93%, specificity of 75% and sensitivity of 92.8%. We obtained an average 6.17 ± 1.66 nm red shift for IDC with respect to normal tissue. The red shift and maximum fluorescence intensity indicates p < 0.01. These results described here are supported by histopathological examination of the same sample.
Conclusion
In the present manuscript, simultaneous fluorescence‐based imaging and spectroscopy is accomplished for the classification of IDC tissues and breast cancer margin detection.
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