Laser speckle imaging has been widely used for in-vivo visualization of blood perfusion in biological tissues. However, existing laser speckle imaging techniques suffer from limited quantification accuracy and spatial resolution. Here we report a novel design and implementation of a powerful laser speckle imaging platform to solve the two critical limitations. The core technique of our platform is a combination of line scan confocal microscopy with laser speckle autocorrelation imaging, which is termed Line Scan Laser Speckle Autocorrelation Imaging (LS-LSAI). The technical advantages of LS-LSAI include high spatial resolution (~4.4 μm) for visualizing and quantifying blood flow in microvessels, as well as video-rate imaging speed for tracing dynamic flow.