In this chapter, we present the optical diagnosis of normal and dengue viral-infected human blood using Raman, Polarimetric, Transmission, and Fluorescence Spectroscopic techniques. The possibility of using light in diagnosis and treating illness has been known for thousands of years. The properties of light and lasers provided many modern applications at home, in industry, and in the field of medicine. Laser use in the field of medicine is large and steadily growing. This growth is based on the versatility of laser light. Efficient and accurate diagnosis of dengue is of primary importance for clinical care. A range of laboratory diagnostic methods has been developed to support patient management and disease control. The choice of diagnostic method depends on the purpose for which the testing is done, the type of laboratory facilities and technical expertise available, costs, and the time of sample collection. The dengue viral infection is mostly diagnosed through laboratory tests; these tests include detection of the virus, virus antigen, anti-dengue virus antibody, complement fixation test, neutralization tests, and detection of virus nucleic acid. As dengue infection most rapidly increases in different regions, early diagnostic confirmation of dengue infection in patients allows for timely clinical intervention, etiological investigation, and disease control. Hence, diagnosis of dengue disease during the acute phase should be a priority and is a public health concern. Lasers and optics have many applications in medical sciences; diagnosis and treatment of diseases with lasers and light are latest and noninvasive techniques. Development of light-based apparatus has evolved into tools for improved diagnosis and treatment modalities in medical sciences. The methods of the laser spectroscopy make it possible to obtain direct information regarding the structure and dynamics of the functional groups of biomolecules. Development of new light sources, optics, and diode laser of different wavelengths makes them attractive for spectroscopy of biological molecules. In our study, more than 600 dengue viral-infected blood or blood sera samples and 25 non-dengue healthy blood samples were analyzed using four different optical methodologies. In the first study, Raman spectrum peaks for normal samples observed at 1527, 1170, and 1021 cm −1 show the presence of different biological materials, including lipids, carbohydrates, skeletal CC stretch of acyl chains, and