Keywords : Digital Holographic Microscopy
journal of kerbala university,
Volume 10, Issue 0, Pages 75-86
In-line DHM was designed, holograms recorded and the numerical reconstructions were made using an algorithm based on the diffraction theory in Fresnel approximation using Fourier Transformation. Using double Nd:YAG laser (λ = 532 nm) output (approximately 30 mW), and CCD camera used with 1024 X 1024 pixels , microscopic objective with 0.87 NA was used for magniﬁcation, obtained images resolution about 0.31µm.we observed there are Inverse relationship between the numerical aperture and the resolution .
for applying this new microscope we used it for a biological sample -red blood cells- that took it from different diseases (e.g. Healthy Person, G6PD, Thalassemia Major ,SCD ,Hereditary Spherocytosis) and making a blood films from it , We present the experimental image result, their 3D image, cross section profiles and histogram. these offer insight into the nature of morphological abnormalities used to identify various disorders, to compute important parameters for each cells have morphological changes (Normal Cell , Anchitocyte , Bite Cell, Elliptocyte, Macrocyte , Spherocyte , Sickle Cell , Stomatocyte , Target Cell , Tear Drop Cell )in this easy way this parameters like Diameter ,Thickness ,Refractive Index , Optical Path Length, Surface Area , Volume ,Spherocity Index.
We found that the diameter of the cells limit between (9.9 µm for macrocyte -3.9 µm for sickle cell) and the normal cell was 7.4 µm , and the thickness of the cells limit between (2.249 µm for Tear-drop cell - 4.233 µm for Spherocyte) and the normal cell was 2.646 µm. refractive index values of 1.18 to 2.22 were and 2.64 for normal cell. The surface areas of individual erythrocytes were measured range from 251.1µm2 for macrocyte to70.8µm2 for sickle cell and the normal cell was 147.70µm2 ,and The volumes of individual erythrocytes were calculated range from 239.25µm3 for macrocyte to 45.8µm3 for sickle cell and the normal cell was 113.98 µm3