Low back pain is the leading cause of disability in the world, with a significant socio-economic impact. Deep dry needling is effective in the treatment of this pain, and it is one of the techniques preferred by physiotherapists. In this field, the use of ultrasound provides information of interest such as length, thickness, diameter, cross-sectional area, or muscle volume, among others. Objective: To find out whether the tissue changes (thickness, histogram, and contraction rate) that occur in the lumbar multifidus after application of the deep dry needle are related to changes in the pain and the disability of the patient. Design: Randomized, double-blind, parallel-group clinical trial. Setting: University of Alcalá, Department of Physiotherapy. Subjects: 21 voluntary patients (women and men) with non-specific low-back pain aged 18–65 years. Intervention: Patients were randomly divided into two groups. One group received dry needling and the other group a dry needling placebo. Initial post-needling and one week post-needling assessments were performed by a therapist blinded to the intervention. Variables: Lumbar multifidus thickness measured by RUSI, contraction time measured by M-mode, histograms measured by image analysis, muscle area, pain measured by VAS, pressure pain threshold measured by pressure algometer, and disability measured by Roland–Morris questionnaire. Conclusions: The contraction speed, resting thickness, and pain demonstrated significant differences within each group, but not between groups. There were significant differences in contraction ratio and in PPT between groups. There was excellent intra-examiner reliability in image collection for histogram analysis. Histogram analysis showed no significant differences between groups and measurements, neither for the parameters nor for the parameters combined with the outcome variables. A robust method for the image texture analyses in future histogram muscle analyses has been performed.