There are Several technological revolutions underway which apply to much of biology like Structural imaging, Functional imaging, Patterned photo-stimulation and Super-resolution techniques which is can use for recording activity from many parts of single neuron in vivo or many neurons in vivo. 2-Photon Microscopy and Linked Optical Methods for Imaging Neurons can give Imaging deep within non-transparent living tissue such as brain….Two-photon fluorescence microscopy allows three-dimensional imaging of biological specimens in vivo. Compared with confocal microscopy, it offers the advantages of deeper tissue diffusion and less photo damage but has the disadvantage of slightly lower resolution. Two-photon microscopy is expected to have an impact in areas such as physiology, neurobiology, embryology and
tissue engineering, for which imaging of extremely scattering tissue is required. I am trying strive to meet the challenge offered by the chance to ask questions about the workings of as cell that we never thought we could answer, we need to be aware that the new technologies are still evolving. The current limitations of each technique need to be measured when identical them to specific biological requests. In this review, we briefly describe the principles of super-resolution optical microscopy and focus on comparing the features of each technique that are significant for their use in learning nanosensing in the cellular microenvironment.