Bi

Towards high-throughput in-situ structural biology

Date
Feb 3, 2020
Time
11:00 AM - 12:00 PM
Speaker
Alex de Marco
Affiliation
Monash University, Dept. of Biochemistry and Molecular Biology, ARC Centre of Excellence in Advanced Molecular Imaging, Clayton, Australia
Language
en
Main Topic
Biologie
Host
Gaia Pigino
Description
Cryo-transmission electron tomography (cryoET) in association with cryo-focused ion beam (cryoFIB) milling enables structural biology studies to be performed directly within the cellular environment. Cryo-preserved cells are milled and a lamella with a thickness of 200-300 nm provides an electron transparent window suitable for cryoET imaging. CryoFIB milling is an effective method, but it is a tedious and time-consuming process, which typically results in ~10 lamellae per day. Here, we introduce an automated method to reproducibly prepare cryo-lamellae on a grid and reduce the amount of human supervision throughout the process. We tested the routine on cryo-preserved Saccharomyces cerevisiae and demonstrate that it is possible to increase the throughput and achieve a rate of 5 lamellae/hour without the need to supervise the preparation. We demonstrate that the quality of the lamellae is consistent throughout the preparation and their compatibility with cryoET analyses. This method in association with the integration of a cryo-light microscope inside the cryoFIB and with the optimization of the ion source leads to the possibility to prepare efficiently targeted lamellae in tissue.

Last modified: Feb 4, 2020, 12:06:26 AM

Location

Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG Galleria)Pfotenhauerstraße10801307Dresden
Phone
+49 351 210-0
Fax
+49 351 210-2000
E-Mail
MPI-CBG
Homepage
http://www.mpi-cbg.de

Organizer

Max Planck Institute of Molecular Cell Biology and GeneticsPfotenhauerstraße10801307Dresden
Phone
+49 351 210-0
Fax
+49 351 210-2000
E-Mail
MPI-CBG
Homepage
http://www.mpi-cbg.de
Scan this code with your smartphone and get directly this event in your calendar. Increase the image size by clicking on the QR-Code if you have problems to scan it.
  • BiBiology
  • ChChemistry
  • CiCivil Eng., Architecture
  • CoComputer Science
  • EcEconomics
  • ElElectrical and Computer Eng.
  • EnEnvironmental Sciences
  • LaLaw
  • CuLinguistics, Literature and Culture
  • MtMaterials
  • MaMathematics
  • McMechanical Engineering
  • MeMedicine
  • PhPhysics
  • PsPsychology
  • SoSociety, Philosophy, Education
  • SpSpin-off/Transfer
  • TrTraffic
  • TgTraining
  • WlWelcome