We investigate microstructures on 316L stainless steel surface induced through using femtosecond laser with central wavelength of 800 nm and pulse duration of 130 fs at a repetition rate of 1 kHz in high vacuum. The single-pulse threshold of 316L stainless steel is obtained by experiment. The multi-pulse laser energy accumulation is experimentally studied with the laser fluences higher and lower than the single-pulse threshold, respectively. Experimental results show that with the laser fluence increasing, laser-induced periodic surface structures (LIPSSs) on the submicron scale, ripples and cone-shaped spikes on the micron scale are fabricated with single-pulse energy of 100 μJ which is higher than the threshold. And LIPSS on the submicron scale and ripples on the micron scale can also be obtained by multiple pulses with single-pulse energy of 20 μJ which is lower than the threshold.