“…CryoCLEM approaches (Lucić et al, 2007;Sartori et al, 2007;Plitzko et al, 2009;Hampton et al, 2017), including those involving cryoVEM methods (Vidavsky et al, 2016) such as cryoSBF-SEM (Hoffman et al, 2020) and cryoFIB-SEM (Gorelick et al, 2019), are not yet routine but constitute an actively developing field given their capacity to highlight molecular identities during ultrastructural examination of cells and tissues without chemical fixation and staining artifacts (Bharat and Kukulski, 2019). Increasing efforts to automate these techniques will help to make them routine for many specimens in the near future (Klumpe et al, 2021;Yang et al, 2021;Weiner et al, 2022), including bacteria and other pathogens (Liedtke et al, 2022), particularly as ML strategies are incorporated into many steps of the workflows (Seifert et al, 2020). Recent implementations have demonstrated cryoCLEM of entire cells using cryoSBF-SEM followed by cryoFIB-SEM lamellae milling and cryoET of targeted areas of interest (Wu et al, 2020), as well as for tissues using the 'lift-out' technique (Schaffer et al, 2019;Kuba et al, 2021) and entire organisms using 'serial lift-out' (Schiøtz et al, 2023).…”