Hydrogen assisted cracking (HAC) is a common type of failure mechanism that can affect a wide range of metals and alloys. Experimental studies of HAC are cumbersome due to various intrinsic and extrinsic parameters and factors (associated with stress, hydrogen and the materials microstructure) contributing to the hydrogen crack kinetics. The microstructure of many materials consists of diverse constituents with characteristic features and mechanical properties which only occur in very small material volumes. The only way to differentiate the effect of these individual constituents on the hydrogen crack kinetics is to miniaturise the testing procedures. In this paper we present a new experimental approach to investigate hydrogen assisted crack growth in a microstructural constituent, i.e. acicular ferrite. For this purpose, sharply notched micro-cantilevers were fabricated with a Focus Ion Beam within this selected microscopic region. Acicular ferrite can be found in many ferrous alloys including ferritic weld metal and has specific features that control its intrinsic susceptibility to HAC. These features were characterised via Electron Backscatter Diffraction and the specimens were subsequently loaded under uncharged and hydrogen charged conditions with a nano-indenter. The outcomes of the testing, demonstrated that the threshold stress intensity factor, Kth, to initiate crack propagation in acicular ferrite ranges between 1.56 MPa m1/2 and 4.36 MPa m1/2. This range is significantly below the values of Kth reported for various ferrous alloys in standard macro-tests. This finding indicates that the mechanisms and resistance to HAC at micro-scale could be very different than at the macro-scale as not all fracture toughening mechanisms may be activated at this scale level.