The invention relates to a three-dimension
microscopic observation method of
plant leaf
stoma in vivo. The method comprises the following steps that: firstly,
optical amplifier components are connected with a focusing guide rail and a varifocus lens is connected with a CCD
image acquisition sensor; the focusing guide rail is connected with a connection plate and the connection plate is connected with a horizontal
fine tuning guide rail; then, a measured
plant is placed on a measured-object load-bearing platform and a observation area of a measured leaf is fixed; the CCD
image acquisition sensor is connected with an upper computer so as to real-timely collecting and observing leaf stomata through
image acquisition software; finally, an image of a leaf surface can be acquired through automatically focusing or manually focusing and a three dimensional image of leaf surface
stoma states can be obtained after performing image denosing,
curve smoothing and
curve fitting on collected data. By using the method of the invention, when the
plant leaves are not separated with the plant body,
stoma state images can be continuously collected, and normal
plant growth state can be two-dimensionally observed or leaf stoma change status under a specific experiment state can be two-dimensionally observed. Therefore, maximum information of the leaf stomata can be guaranteed.