AbstractMost applications of multi-photon microscopy (MPM) in biology and medicine focus on high-resolution imaging of physiology, morphology and cell–cell interactions in intact tissues or live animals. In this paper, our aim is to assess whether MPM has the ability to provide comparable information for stained histology slides and to identify inflammatory fibroid polyp (IFP) tissue sections without H&E staining.A tissue section without H&E staining was examined using a multi-photon microscope

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... consisting of a mode-locked Ti: sapphire femtosecond laser (Mira 900-F; Coherent, Inc.) and a commercial laser scanning microscope (LSM 510 META; Carl Zeiss, Inc.) with a Plan-Neofluar 40×/0.75 objective. Large area images and a series of optical sectioning images were obtained using an optional fine-focusing stage (HRZ 200; Carl Zeiss, Inc.).MPM images showed the main characteristics of IFP that corresponded well with H&E microscopic results, such as a typical onion skin arrangement of reticular fibers around thin-walled blood vessels, and short spindle cells intermixed with abundant inflammatory cells. A series of optical sectioning images displayed morphological structure of tissue sections at different depths.MPM has the ability to provide comparable results to H&E staining. Its capability of optical sectioning can present the morphological structure of different optical sectioning planes, making MPM an alternative for those cases where the sample is very thick or cannot be stained.