Led EVs into mice showed their transport into lymph nodes and internalization by antigen-presenting cells, specifically these expressing CD11b. Summary/Conclusion: In conclusion, glycan evaluation of EVs using a lectin array program is usually a uncomplicated and valid tool for the EV standardization and EV-cell interaction. Reference: [1] Shimoda A, et al. Biochem Biophys Res Commun. 2017;491:70107.Methods: Cryo-immobilization of MMP-17 Proteins Storage & Stability bacteria and MVs by HPF-FS and TEM; cryo-TEM of plunge-frozen whole bacteria and MVs; encapsulation of DNA inside the MVs by TEM right after gold DNA immunolabelling. Benefits: The use of these methods revealed some interesting findings. Initial, the structural evaluation of your extracellular matter created by many Gram-negative Antarctic bacteria following HPF-FS TEM permitted us to establish its complexity, appearing as a netlike mesh containing substantial numbers of MVs. The release of MVs through bulging and “pinching off” from the outer membrane was confirmed. On top of that, we demonstrated a new model of vesiculation in both environmental and pathogenic bacteria that results in the formation of a distinct variety of outer membrane vesicle with a double-bilayer structure, which encapsulates DNA and thus could be involved in DNA transfer. Furthermore, we detected that the introduction of mutations in bacterial strains to Interferon-Stimulated Gene 15 (ISG15) Proteins web induce hypervesiculating phenotypes results in alterations in MV composition and in their potential to interact with host cells, which is usually explained by considerable modifications in MVs structure and this might have a significant impact on MV functionality. Summary/Conclusion: This study exposes the need for conducting a detailed structural analysis by high-resolution TEM techniques when operating with MVs. This evaluation must be mandatory as a way to assure the good study practice in MV investigation field, especially if they’re intended to be made use of for therapeutic purposes. Funding: This study was funded by Government of Spain (CTQ201459632-R). CPC received the fellowship APIF2015 from the UB, and NB BES2015-074582 in the Government of Spain.PS09.Enhancing accuracy of clinical predictions on shifted microflow cytometry information with signal standardizationRobert J. Paproski1; Desmond Pink1; Renjith Pillai2; Catalina Vasquez2; John D. LewisUniversity of Alberta, Edmonton, Canada; CanadaNanostics Inc, Edmonton,PS09.TEM and Cryo-TEM microscopy as a tool to elucidate prokaryotic membrane vesicle structure Carla Perez-Cruz1; Nicolas Baeza1; Carmen Lopez-Iglesias2; Elena Mercade1 Department of Biology, Well being and Atmosphere, University of Barcelona, Barcelona, Spain; 2The Maastricht Multimodal Molecular Imaging institute, Maastricht University, Maastricht, The NetherlandsBackground: There is a need to have to characterize the structure of membrane vesicles (MVs). In most published studies, MVs morphology and integrity is revealed by transmission electron microscopy (TEM) micrographs from negatively stained MVs, however the resolution of this strategy just isn’t adequate. TEM observation of specimens cryoimmobilized by high pressure freezing (HPF) followed by freeze substitution (FS) and sectioning, with each other with cryo-TEM observation of frozen-hydrated specimens, allow the visualization of biological samples close to their native state, enabling us to refine our information of bacterial structures such us MVs.Background: We have developed a state-of-the-art XGBoost-based algorithm for predicting clinical outcomes from microflow cytometry information which considerably ou.