Cryogenic Transmission Electron Microscopy
Cryogenic Transmission Electron Microscopy-IoE RIFC
Make
Thermofischer Krios G4
Model
Thermofischer Krios G4
Facility Status
Working
Facility Management Division
Institute Central Research Facilities (ICRF)

Category

  • Microscopy and Imaging » Electron Microscopy

Booking Details

Booking available for
Internal and External Both
Available Mode for Use
Imaging techniques for both cryogenic and negative-stained Transmission Electron Microscopy (TEM).

Facility Management Team and Location

Facility Manager

Prof. Indradev Samajdar 
Prof. Prasenjit Bhaumik 
Prof. Roop Mallik 
Prof. Pradeepkumar P.I. 
Prof. Shobana Kapoor
Prof. Sarika Mehra 

Prof. Ajit Rajwade
Prof. Sandip Kaledhonkar
Co-convenors
Prof. Ruchi Anand
LAB Email ID
cryoem@ircc.iitb.ac.in
Facility Location
Lab no. 310, Cryogenic Transmission Electron Microscopy Facility (120kV & 300kV CryoEM), Ground Floor, SAIF/CRNTS Department, IIT Bombay.
Lab Phone No
022 2576 3358

Facility Features, Working Principle and Specifications

Features Working Principle

The 300 keV Krios G4 cryo-electron microscope by Thermofisher is an advanced tool for atomic resolution cryo-EM analysis, boasting enhanced productivity. It comes equipped with a BioContinuum GATAN K3 detector and energy filter from Amtek-Gatan, facilitating both single-particle and tomography studies. Complementing the microscope are all the necessary accessories for frozen-hydrated sample preparation for single particle reconstruction and cryotomography. Moreover, the facility houses a 120-keV screening microscope and various accessories like the Vitrobot, Cryo-Ultramicrotome, and High-Pressure Freezer, which aid in sample preparation and method development. This comprehensive setup enables the facility to capture high-resolution data on biomolecules such as proteins, protein complexes, viruses, and nanoparticles, providing detailed insights into cell and tissue sections.

Cryo EM is an advanced imaging technique used to elucidate the three-dimensional (3D) structure of biological molecules and complexes at near-atomic resolution. In this technique, the sample is rapidly frozen to temperatures below -150 °C, trapping it in vitreous ice.

Body Specification

Microscope: Krios G4
Imaging: Utilizing an Extreme High-Brightness Field Emission Gun (X-FEG) and offering a flexible accelerating voltage range from 80 to 300 kV, complete with automatic condenser, objective, and SA apertures to facilitate multiple image acquisitions per grid hole.
Detector: Equipped with both BioContinuum GATAN K3 detector and Ceta D detectors.
Energy Filter: Incorporating the Biocontinuum K3 energy filter.
Software for Data Collection and Screening: Utilizing EPU for streamlined data collection and screening processes.

Instructions for Registration, Sample Preparation, User Instructions, Precautionary Measures and Charges

Instructions for Registration

Internal users are required to register online, while external users are also instructed to register online. Appointments will be allocated based on the queue and communicated via email. New users are encouraged to reach out to the CryoEM lab before initiating the registration process.

Instruction for Sample Preparation

Prior to securing a slot for the Krios 300kV microscope, users must first examine the sample on a negatively stained grid, followed by screening on a cryo-conditioned grid using the Talos 120kV microscope. This data should be submitted at the time of slot booking.

User Instructions and Precautionary Measures
  • Online registration is required for users.
  • Pre-registration is mandatory for external users.
  • Appointments will be scheduled based on the queue and communicated via email.
  • New users are advised to reach out to the CryoEM lab prior to registration.
  • Users must provide at least 24 hour’s notice to the CryoEM lab if they wish to cancel or postpone their slot.

Applications

  • Biological sciences.
  • a) Well-suited for single-particle analysis, electron tomography, and micro electron diffraction.
  • b) Its Cryo functionality renders it versatile for applications in medicine, biology, pharmacy, and the study of unknown materials.
  • c) Capable of performing 3D analysis and imaging.
  • d) Enables analysis at the molecular level.
  • e) Facilitates structural-based drug design.

Sample Details

SOP, Lab Policies and Other Details

Publications

Publications
NA