An investigation into orientation preferences shown by actin fibres within ex-situ actin as imaged by Atomic Force Microscopy (AFM) is described. Actin is a primary cytoskeletal component and is believed to play a vital role in cell structure. Actin structure images measured by AFM were analysed using automated pre-processing steps. These steps were identical for the production of an initial binary image, which was then processed by both Hough transformation and thinning. Results obtained question the validity of using the Hough transform approach, as bias could not easily be eliminated, and hence the Hough transform method was deemed to be unsuitable for this application and instead the thinning technique was used to identify and locate actin orientation within the AFM images. The results show that polymerised ex-situ actin fibres appears to display a bimodal distribution of orientation, with a 90 degree separation, with a significant co-efficient of bimodality of 0.656.