Transforming cryo-lift-out one section at a time

Understanding the complexity of cells – their components, processes, and dynamics – requires imaging techniques capable of capturing biological organization in situ and across scales. Advances in cryo-electron tomography (cryo-ET), enabled by focused ion beam (FIB) milling, have positioned this technique as a powerful tool for visualizing cells at molecular resolution. Extending cryo-ET to multicellular specimens such as organoids or tissues, however, requires sample preparation strategies that can access areas deep within complex specimens. Cryo-lift-out has emerged as a key sample preparation technique to address this challenge. Here, we review recent advances in cryo-lift-out to image cellular processes, discuss the remaining technical challenges, and outline our perspectives on how continued methodological development may enable more holistic in situ studies in the future.