IADR Abstract Archives
Lateral Shearing Digital Holographic Microscope for Imaging of Oral Pathology
Objectives: Oral and maxillofacial pathology relies on staining and labeling of biological/biopsy specimens for diagnosis and treatment planning. Digital holographic microscopy is an upcoming label-free quantitative technique to observe biological tissue specimens. This study was conducted to design and fabricate a novel wide field of view common-path lateral shearing Digital Holographic quantitative phase microscope (LS-DHM) for imaging and quantifying the morphology and dynamics of hyperplasia, dysplasia and cancerous oral tissue specimens.
Methods: We designed LS-DHM setup consisting of coherent light source (LASER), microscopic objective for magnification, wedged glass plate and detector viz., charged couple device (CCD) or complementary metal-oxide-semiconductor (CMOS) camera. LS-DHM setup was calibrated by recording holograms of polystyrene microspheres, immersed in microscope oil. Spatial and temporal phase stability of microscope was also evaluated. 103, 94 and 121 fibrous hyperplasia, epithelial hyperplasia with mild to moderate dysplasia and well-differentiated squamous cell carcinoma specimens, respectively, obtained from human patients referred for oral biopsy, after informed consent, were subjected to routine hematoxylin and eosin staining after fixing whereas fixed unstained samples were observed under the novel LS-DHM.
Results: LS-DHM calibration revealed mean thickness value of 10.11μm ± 0.19μm for polystyrene microspheres, which was quite close to manufacturer specified value of 10 μm, indicating its 3D imaging capability. Spatial phase stability of microscope in Phase (rad) measured using CCD array was 0.042 ± 0.0047 whereas temporal phase stability was 0.00821 ± 0.00091. High quality holographic images of fibrous hyperplasia, epithelial hyperplasia with mild to moderate dysplasia and well-differentiated squamous cell carcinoma specimens exhibited significant changes in cellular morphology and dimensions, which correlated with histological findings in stained specimens
Conclusions: The novel LS-DHM exhibited excellent 3D imaging capability; was spatially and temporally stable and was able to demonstrate true holographic images of oral lesions, which correlated with histological findings in stained specimens
Methods: We designed LS-DHM setup consisting of coherent light source (LASER), microscopic objective for magnification, wedged glass plate and detector viz., charged couple device (CCD) or complementary metal-oxide-semiconductor (CMOS) camera. LS-DHM setup was calibrated by recording holograms of polystyrene microspheres, immersed in microscope oil. Spatial and temporal phase stability of microscope was also evaluated. 103, 94 and 121 fibrous hyperplasia, epithelial hyperplasia with mild to moderate dysplasia and well-differentiated squamous cell carcinoma specimens, respectively, obtained from human patients referred for oral biopsy, after informed consent, were subjected to routine hematoxylin and eosin staining after fixing whereas fixed unstained samples were observed under the novel LS-DHM.
Results: LS-DHM calibration revealed mean thickness value of 10.11μm ± 0.19μm for polystyrene microspheres, which was quite close to manufacturer specified value of 10 μm, indicating its 3D imaging capability. Spatial phase stability of microscope in Phase (rad) measured using CCD array was 0.042 ± 0.0047 whereas temporal phase stability was 0.00821 ± 0.00091. High quality holographic images of fibrous hyperplasia, epithelial hyperplasia with mild to moderate dysplasia and well-differentiated squamous cell carcinoma specimens exhibited significant changes in cellular morphology and dimensions, which correlated with histological findings in stained specimens
Conclusions: The novel LS-DHM exhibited excellent 3D imaging capability; was spatially and temporally stable and was able to demonstrate true holographic images of oral lesions, which correlated with histological findings in stained specimens