In order to obtain high yield of astaxanthin, a high-value compound with ultrastrong antioxidant capacity, it is necessary to identify the growth characteristics (biomass, morphology, and size) of Haematococcus pluvialis. The current detection methods have the disadvantages of labor-consuming operation or complicated measurement system. It is an urgent need to explore a simple and cost-effective method for the detection of H. pluvialis with large size distribution during its growth period. In this work, a digital in-line holographic flow cytometry using a linear array sensor is proposed to measure the growth characteristics of H. pluvialis in a two-dimensional (2-D) hydrodynamic focusing microfluidic chip. Based on the modified angular spectrum method, the distorting holograms caused by the asynchrony of sample flow velocity and acquisition speed of the linear array sensor were rectified and reconstructed. In addition, the depth-of-focus of the imaging system were digitally extended to cover the entire depth of the microfluidic channel for optimized imaging quality. We have utilized the proposed method to statistically investigate the biomass, morphology and size of H. pluvialis under different culture conditions and growth durations.