The invention relates to a locating and guiding device in a minimally invasive surgery, which comprises a main frame and a handheld component, wherein the handheld component comprises a housing, a projecting device, a pick-up device, a front imaging processing device and a handheld component control device, wherein the projecting device is used for emitting an illuminating beam the pick-up device is used for receiving the imaging of a returned beam the front imaging processing device is designed with the pick-up device in an integrated manner so as to carry out imaging process on a returned optical signal and the handheld component control device is provided with a control button and a control state indication lamp and is electrically connected with the projecting device so as to control the switch of light sources, and is used for adjusting the light intensity by changing the power of current output. The projecting device comprises the first light source for specifically organizing the imaging of an absorption spectrum and the second light source for stimulating the imaging of a fluorescent substance the pick-up device and an image pre-processing device are provided with narrow bandpass filters and the device is such constructed that imaging is carried out on the absorption spectrum with specific organization and imaging is carried out on the exciting light of the fluorescent substance simultaneously or individually by utilizing the same pick-up image and image preprocessing device. Through the device, imaging can be carried out on blood vessels and lymph simultaneously or individually without light splitting design.
