Electron microscopy stands as a cornerstone in unraveling the intricate dynamics of viral infections, with its high-resolution capabilities offering invaluable insights into the interactions between viruses and the infected cells. Here, we present a comprehensive methodology designed to explore the three-dimensional interactions specifically between tick-borne encephalitis virus (TBEV) and host cells. This approach allows to study all stages of viral lifecycle, including replication, budding, maturation, and host cell defense mechanisms. The methodology encompasses a range of techniques, commencing with sample preparation using high-pressure freezing, followed by freeze substitution, epoxy embedding, and ultrathin sectioning. Subsequently, we employ electron tomography in conjunction with image processing and analysis techniques to unravel the intricate nuances of TBEV-host cell interactions.

• Klíčová slova
• Electron tomography, Tick-borne encephalitis virus, Transmission electron microscopy,
• MeSH
• buněčné linie MeSH
• interakce hostitele a patogenu * MeSH
• klíšťová encefalitida * virologie   MeSH
• lidé MeSH
• mrazová substituce MeSH
• replikace viru MeSH
• tomografie elektronová * metody   MeSH
• viry klíšťové encefalitidy * fyziologie   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Vector-borne diseases constitute 17% of all infectious diseases in the world; among the blood-feeding arthropods, ticks transmit the highest number of pathogens. Understanding the interactions between the tick vector, the mammalian host and the pathogens circulating between them is the basis for the successful development of vaccines against ticks or the tick-transmitted pathogens as well as for the development of specific treatments against tick-borne infections. A lot of effort has been put into transcriptomic and proteomic analyses; however, the protein-carbohydrate interactions and the overall glycobiology of ticks and tick-borne pathogens has not been given the importance or priority deserved. Novel (bio)analytical techniques and their availability have immensely increased the possibilities in glycobiology research and thus novel information in the glycobiology of ticks and tick-borne pathogens is being generated at a faster pace each year. This review brings a comprehensive summary of the knowledge on both the glycosylated proteins and the glycan-binding proteins of the ticks as well as the tick-transmitted pathogens, with emphasis on the interactions allowing the infection of both the ticks and the hosts by various bacteria and tick-borne encephalitis virus.

• Klíčová slova
• Anaplasma, Borrelia, Carbohydrate-binding, Glycan, Glycobiology, Host, Lectin, Pathogen, TBEV, Tick,
• MeSH
• Anaplasma patogenita   MeSH
• Borrelia patogenita   MeSH
• glykomika metody   MeSH
• glykosylace MeSH
• interakce hostitele a patogenu fyziologie   MeSH
• klíště mikrobiologie   fyziologie   virologie   MeSH
• lektiny metabolismus   MeSH
• nemoci přenášené klíšťaty patofyziologie   MeSH
• polysacharidy metabolismus   MeSH
• proteomika MeSH
• sacharidy fyziologie   MeSH
• viry klíšťové encefalitidy patogenita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• lektiny MeSH
• polysacharidy MeSH
• sacharidy MeSH

To understand the response of the Lyme disease spirochete Borrelia burgdorferi exposed to stress conditions and assess the viability of this spirochete, we used a correlative cryo-fluorescence and cryo-scanning microscopy approach. This approach enables simple exposition of bacteria to various experimental conditions that can be stopped at certain time intervals by cryo-immobilization, examination of cell viability without necessity to maintain suitable culture conditions during viability assays, and visualization of structures in their native state at high magnification. We focused on rare and transient events e.g., the formation of round bodies and the presence of membranous blebs in spirochetes exposed to culture medium, host sera either without or with the bacteriolytic effect and water. We described all crucial steps of the workflow, particularly the influence of freeze-etching and accelerating voltage on the visualization of topography. With the help of newly designed cryo-transport device, we achieved greater reproducibility.

• Klíčová slova
• Borrelia burgdorferi, Lyme disease, cryo-fluorescence, cryo-scanning electron microscopy, pleomorphism, round body, viability staining,
• Publikační typ
• časopisecké články MeSH