Objective To establish a method  of in vivo microvascular imaging and blood flow calculation with better continuous imaging quality.   Methods Anesthetized mice with dye injection through tail vein were placed in the center of a 37 ℃ hot plate holder. The stripped tissues were placed in a self-made low edge confocal dish and immersed in normal saline. The exposed tissues were pressed with self-made circular metal pads of different weights and sizes. The microvascular blood flow in the liver and hind limb of pregnant mice (n=3) and normal female mice (n=3) were measured by the improved device.   Results This method  can accomplish stable and continuous imaging. The blood flow velocity of liver and hind limb of pregnant mice were faster than that of normal female mice.  Conclusion This method  can be used for three-dimensional imaging of microvessels and detection of blood flow velocity in organs.