Operating F20/F30 with SerialEM

Chen Xu

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This is a quick check list for the Tecnai F20 or Tecnai F30 users who already have basic knowledge of Tecnai FEG microscope. I list here the steps that I would perform if I am on the scope with cryo sample and plan to collect single particle data using SerialEM.

A pdf version of this document is also available.

Table of Contents
1 Check E-logbook to see if there have been any problems
2 Login to Tecnai Computer
3 Check vacuum status
4 Check settings for H.T., Gun Lens and Extracting Voltage
5 Cool down the scope
6 Check CryoBox Position
7 Put in a single tilt holder with a standard grid
8 Eucentric height, Eucentric focus and Pivot Points
9 Check coma-free and beam conditions for high-res imaging
10 Start up CCD or 3rd party software interfaces
11 Set Bias for 4-port CCD
12 Prepare Gain Reference Files
13 Setup Low-Dose in SerialEM
14 Cryo Holder In
15 Stop LN2 bubbling in the holder dewar
16 Eucentricity, without wobbling the stage if possible.
17 Obtain Full Grid Overview
18 Resume Low-Dose Mode
19 Mark Interested Points
20 Check your main Macro
21 Acquire at all points
22 Finishing Up

1 Check E-logbook to see if there have been any problems

It is not a bad idea to see if there is any problem or issue from last user that should have been logged on the E-logbook.