tilt series

Other

65

(4096, 4096)

SIGNED 16 BIT INTEGER

(7.5, 7.5)

HeLa cells were instantly frozen using a high-pressure freezing machine (HPM 010; ABRA Fluid AG, Widnau, Switzerland) and freeze-substituted into Lowicryl HM20 resin (Polysciences Inc., Warrington, PA). As fiducial markers, 15 nm of gold-conjugated Protein A (CMC university Medical Center Utrecht, Utrecht, Netherlands) was absorbed on both sides of 300 nm sections. Sections were post-stained with 2% UA and lead citrate. Single or dual axis tilt series were acquired with a TECNAI TF30 trans-mission EM (TEM; 300 kV; FEI, Hillsboro, OR) equipped with a 4k x 4k Eagle camera (FEI) by using the Serial EM software. Images were recorded over a -60° to 60° tilt range with an angular increment 1°.

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