Due to their ability to carry out both additive and subtractive processing of a wide range of materials in a non-contact manner with superior precision and speed, lasers are a promising tool to create reproducible constructs for cell based assays. Laser direct write technologies – such as our custom-designed matrix assisted pulsed laser evaporation (MAPLE) bioprinting system – combine the power of laser processing with computer aided design to allow us to print as well as ablate cells, cell aggregates and biomaterials with high resolution. Here, we present some interesting applications of the excimer laser driven MAPLE platform for cell printing and ablation, tissue dissection, biomaterial micromachining and 3D microbead fabrication for studying interactions between cells and their microenvironmental cues in vitro. We have printed breast and colon cancer cells, fibroblasts and macrophages in spatially defined patterns on hydrogel substrates and live rat mesenteric tissue to develop physiologically relevant models for cancer cell migration and invasion. We have micromachined spoke and channel patterns into PDMS substrates to analyze competitive cell migration in context of cancer and atherosclerosis. We have also developed a reproducible and customizable wound healing assay using laser ablation of adherent cells. We have fabricated and patterned 3D microbeads encapsulating heterogeneous cell types to elucidate cancer-adipose interactions. Finally, we have dissected user-specified regions of excised mouse colonic tissue to create testable sections. Our work demonstrates the immense versatility of laser based direct write technologies in biology and medicine.