TFS Vitrobot Mark IV

Merit Badge 4

Category: Sample preparation
Sub-category: Cryogenic work

Plunge freezing and instrument certification for Vitrobot Mark IV.

Theoretical background

Knowledge of the instrument

Safety

Specific safety SOPs for this merit badge.

• NCCAT SOP for work with ethane for plunge freezing
For general safety information see the SOPs in the right bar.

Relevant SDS (Safety Data Sheets)

• Airgas-SDS-LN2-001188
• Airgas-SDS-Ethane-001024

Vendor product information

Sample preparation equipment for electron microscopy – CryoEM sample preparation with the Vitrobot System for reproducible vitrification of biological samples

• TFS Vitrobot Mark IV product page.
• TFS Vitrobot Mark IV data sheet.

Theoretical knowledge

Curriculum development partner modules

Use of the instrument
• Getting Started in CryoEM – Unit 2: Sample Preparation [Plunge freezing with a Vitrobot modules]
• CryoVR – Vitrobot module

Center specific guidelines

• NCCAT General Vitrobot Mark IV SOP

Knowledge test

A pass/fail test

The knowledge assessment test is a shared test between centers and may be updated regularly. If a user has earned the merit badge and policies have been updated, then an updated information sheet would have to be reviewed ahead of booking an instrumentation session.

Demonstration

Center staff training

The instrument demonstration should be scheduled with center staff through the NCCAT User Office or your appointed user coordinator.

Practice

Supervised and unsupervised practice by the user

Center staff will assist in arranging dedicated time slots for access to the necessary equipment. Registered users will be issued scheduler accounts through the NCCAT User Office.

Practical test

Two (2) separate center staff observing your proficiency at separate times

Center staff will assist in arranging dedicated time slots for access to the necessary equipment. Registered users will be issued scheduler accounts through the NCCAT User Office.

Additional information

Recertification period

  • Sample preparation merit badges for cryogenic work are valid for ~6mo.
  • Recertification (to maintain active status) would entail demonstrating proper use and care in front of a center staff member.

Furthering your knowledge

General background on plunge freezing