EMD-5627 Experiments and Validation

Cryo-electron tomography of in situ flagellar motor of Borrelia burgdorferi

Subtomogram averaging
Overview of EMD-5627
Sample name: In situ flagellar motor from B. burgdorferi wild type cells
Organism: Borrelia burgdorferi
Related in-frame EM entries: EMD-5628, EMD-5629, EMD-5630, EMD-5631, EMD-5632, EMD-5633
Related EM entries by publication: EMD-5629, EMD-5628, EMD-5630, EMD-5631, EMD-5632, EMD-5633

Map parameters

Minimum density: -18.76
Maximum density: 15.12
Average density: 0
Standard deviation: 1
Recommended contour level: 1.5 (author)

Sample information

Sample name: In situ flagellar motor from B. burgdorferi wild type cells
Cellular component: flagellar motor

Validation information

Experimental information

 

Image processing

Applied symmetry: C1
Software: IMOD, Raptor, Protomo
Number of subtomograms averaged: 999
Number of class averages: 4
Reconstruction protocol: subvolume averaging

Imaging

Session
Microscope model: FEI POLARA 300
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 100.0
Nominal CS (mm): 2
Nominal magnification: 31000.0
Defocus max (nm): 6,000
Defocus min (nm): 4,000
Tilt max (degrees): 65
Tilt min (degrees): -65
Holder model: OTHER
Acquisition
Number of images: 110142

Specimen

Preparation
Specimen state: cell
Buffer: BSK-II liquid medium supplemented with 6% rabbit serum
Specimen support: freshly glow-discharged holey carbon grids
Vitrification
Apparatus: HOMEMADE PLUNGER
Cryogen: ETHANE
Protocol: Bacterial culture was deposited onto freshly glow-discharged, holey carbon grids for 1 min. Grids were blotted with filter paper and then rapidly frozen in liquid ethane, using a homemade gravity-driven plunger apparatus.