The objective of this work was low-cost fabrication of ITO electrodes through a novel laser assisted method. We successfully optimized the cutting of ITO coated glass plates into functional electrodes with a CO2 laser. The electrodes of different shapes and sizes were prepared using two different methods. For the smaller rectangular shaped microelectrodes (smallest widths optimized were 25, 50 and 100 µm wide), direct patterning was used, while in-situ stencil fabrication followed by etching was observed as the best practice for larger circular shaped electrodes (diameter 1.75 mm, 2.41 mm 2 of geometrical surface area). All the electrodes and electrode arrays thus fabricated, demonstrated stable and reproducible signals. Cost-effective, easily fabricated, simple, and handy ITO electrodes we presented here can be treated as any contemporary working electrode in use and as a tool in several fields like sensing, bio-sensing, EWOD, microfluidics, cell culture studies and much more. As a demonstration, we used the electrode arrays for cell culture characterizations using electrochemical impedance spectroscopy (EIS).