Microtubules often form sparse networks in eukaryotic cells which simultaneously contribute to shape maintenance and help establish overall cell layout. It is therefore important to quantify not only how these filaments function individually but also as a coupled network. We have developed a straightforward approach to assemble such networks de novo and we now use it to measure microtubule rigidity within small networks under controlled conditions. Our results suggest that microtubule rigidity increases with the contour length of the filament both for single microtubules and within small microtubule networks. (C) 2020 Elsevier Inc. All rights reserved.