Simulation of light microscope image assists in interpretation and allows comparison of different imaging techniques and their associated parameters. Further, reconstruction of specimen properties from images can in some cases be feasible.Imaging methods can be broadly divided into fluorescent and non-fluorescent techniques, the latterconsisting of brightfield transmission or reflection modes, darkfield or various phase contrast modes. The distinctive feature of these non-fluorescent modes is that they image the material properties of the specimen, including refractive index, absorption coefficient or birefringence. Sometimes the specimen is labelled with a stain, the concentration of which mirrors the concentration of a particularconstituent. In fluorescence microscopy, usually a fluorochrome is used to label particular components or constituents.In non-fluorescent microscopy, images can be simulated using rigorous diffraction methods, such asmodal analysis or coupled wave theory. Such theories take into account multiple scattering, shadowingeffects and also depletion and aberration of the light as it penetrates the object.
Simultaneous measurement of flame temperature and absorption coefficient through LMBC-NNLS and plenoptic imaging techniques
