A computational method for two-dimensional quantitative analysis of standing wave images of red blood cells

Standing wave (SW) microscopy allows for an improvement in the axial resolution that can be obtained in optical microscopy. In SW microscopy two counter propagating waves interfere producing a SW with anti-nodal planes that are separated by λ/2n and a FWHM of λ/4n which is the axial resolution, where λ is the excitation wavelength and n is the refractive index Multi-planar SW microscopy, with the addition of a mirror below the specimen, allows for selective plane contour mapping of one concave surface of a red blood cell (RBC). We report a computational method to extract SW anti-nodal plane and boundaries positions (x,y) and extract each pixel intensity value. By doing so, we can create 2D reconstruction of SW RBC images captured at video rate. In future, by utilising the positional information for the each of the anti-nodal SW planes, we aim to create 3D and 4D reconstruction of the RBCs concave surface. Additionally, applying the extended computational method to SW RBC images we aim to study healthy and diseased erythrocytes and the changes in the concave surface morphology over time.