The fabrication of carbon black/polylactic acid (PLA) electrodes using a 3D printing pen is presented and compared with electrodes obtained by a desktop fused deposition modelling (FDM) 3D printer. The 3D pen was used for the fast production of electrodes in two designs using customized 3D printed parts to act as template and guide the reproducible application of the 3D pen: (i) a single working electrode at the bottom of a 3D-printed cylindrical body and (ii) a three-electrode system on a 3D-printed planar substrate. Both devices were electrochemically characterized using the redox probe [Fe(CN)] via cyclic voltammetry, which presented similar performance to an FDM 3D-printed electrode or a commercial screen-printed carbon electrode (SPE) regarding peak-to-peak separation (ΔEp) and current density. The surface treatment of the carbon black/PLA electrodes fabricated by both 3D pen and FDM 3D-printing procedures provided substantial improvement of the electrochemical activity by removing excess of PLA, which was confirmed by scanning electron microscopic images for electrodes fabricated by both procedures. Structural defects were not inserted after the electrochemical treatment as shown by Raman spectra (i/i), which indicates that the use of 3D pen can replace desktop 3D printers for electrode fabrication. Inter-electrode precision for the best device fabricated using the 3D pen (three-electrode system) was 4% (n = 5) considering current density and anodic peak potential for the redox probe. This device was applied for the detection of 2,4,6-trinitrotoluene (TNT) via square-wave voltammetry of a single-drop of 100 μL placed upon the thee-electrode system, resulting in three reduction peaks commonly verified for TNT on carbon electrodes. Limit of detection of 1.5 μmol L, linear range from 5 to 500 μmol L and RSD lower than 4% for 10 repetitive measurements of 100 μmol L TNT were obtained. The proposed devices can be reused after polishing on sandpaper generating new electrode surfaces, which is an extra advantage over chemically-modified electrochemical sensors applied for TNT detection.