Stimuli-responsive microgel-based etalons are promising optical and bio-sensors. These sensors play a pivotal role in modern healthcare by enabling rapid biomolecule detection, contributing to organ-on-chip applications and early disease diagnosis. This study investigates the suitability of poly(N-isopropylacrylamide) (pNIPAm)-based microgels for inkjet printing, focusing on optimizing their properties for effective deposition. Key parameters, including surface tension, viscosity, and particle size, are characterized to ensure compatibility with inkjet-printing requirements. The addition of surfactants tunes the suspensions’ properties to be in line with the requirements of inkjet printing. Jetting of pNIPAm-based microgels on gold-coated substrates forms a cohesive drop in a range of a few millimeters. Optical and scanning electron microscopy confirm the formation of a uniform microgel layer. The optical reflectance spectroscopy results indicate that inkjet-printed microgel-based etalons can effectively respond to changes in temperature and glucose concentration. In-liquid atomic force microscopy demonstrates the swelling dynamics of the microgels in different glucose concentrations, shedding light on their response dynamics. Our work demonstrates, for the first time, the feasibility of printing microgels in a controlled way, fabricating biocompatible inkjet-printed microgel-based etalon sensors with precise dimensions. The size precision and the sensitive monitoring capabilities of biomolecules hold great promise for in situ and continuous sensing in a wide range of biological and organ-on-chip applications.