TFS Vitrobot Mark IV
Category: Sample preparation
Sub-category: Cryogenic work
Plunge freezing and instrument certification for Vitrobot Mark IV.
You must complete all required safety training at the center you are visiting before hands on training. Coordinate this with the user office or training coordinator.
Relevant Training Modules (to review until you can pass the knowledge test):
- Review the basics of plunge freezing: CryoEM101 – Chapter 2
- Watch the Vitrobot in action will full explanations of instrument use: Getting Started in CryoEM, Unit 2: Sample Preparation [Watch plunge freezing with a Vitrobot modules: 7-20]
- Practice on your own using a virtual instrument: CryoVR – Vitrobot module
SOP for Vitrobot use (for review before demonstration)
Vendor product information for reference
- TFS Vitrobot Mark IV product page (Datasheet available under “Documents”)
The pass/fail knowledge assessment will test understanding of the following concepts:
- Why Ethane is used (and boiling/melting points of ethane/LN2)
- Safety considerations for cryogens/required PPE
- Why formation of vitreous ice is required
- How ice thickness affects data quality
- The reasons for humidity and temperature control
- ID parts of the instrument
Here is the current quiz version:
A workflow demonstration should be scheduled with center staff. The demonstration will cover all points of the SOP. You must have passed the knowledge quiz and reviewed the SOP before the demonstration.
Each center has additional specific Safety, Setup, and Shutdown procedures. Below, you will find those protocols for each center.
While COVID-19 restrictions are in place, practicing freezing can be difficult to come by. In lieu of on-site practice, utilize the latest Virtual Reality technology and practice vitrification from the comfort of your own home with CryoVR from Purdue. No need to worry about lab safety!
When you’re ready to travel for your training, schedule your supervised practice with the user office or appointed user coordinator. This can be scheduled for the same session as the demonstration. During supervised practice, center staff will be present for the full duration of the session to provide feedback and answer questions with the goal that you can pass the practical test for independence with no assistance. The number of supervised practice sessions will be tailored to the user, and if necessary, more than one session can be scheduled before taking the practical test.
Center staff will assist in arranging time for access to the necessary equipment and testing for independence. The practical test can not be scheduled during the same session as supervised practice.
A Vitrobot independence checklist will be used by the certifying staff to test for proper use of the instrument with a pass/fail outcome. All steps on the checklist must be performed to pass. You may use any instructions normally posted as a guide during the session.
There is also an evaluator version of the checklist for more extensive training.
- Sample preparation merit badges for cryogenic work are valid for 6 months.
- Recertification (to maintain active status) requires passing only the practical test again with one center staff member. If supervised training is needed to pass the practical test, this can be arranged.