EIPOD fellowship under Marie Sklodowska-Curie Actions COFUND
664726
European Union
European Research Council (ERC)
760067
European Union
European Union (EU)
871037
European Union
引用
ジャーナル: Elife / 年: 2021 タイトル: A modular platform for automated cryo-FIB workflows. 著者: Sven Klumpe / Herman Kh Fung / Sara K Goetz / Ievgeniia Zagoriy / Bernhard Hampoelz / Xiaojie Zhang / Philipp S Erdmann / Janina Baumbach / Christoph W Müller / Martin Beck / Jürgen M Plitzko / Julia Mahamid / 要旨: Lamella micromachining by focused ion beam milling at cryogenic temperature (cryo-FIB) has matured into a preparation method widely used for cellular cryo-electron tomography. Due to the limited ...Lamella micromachining by focused ion beam milling at cryogenic temperature (cryo-FIB) has matured into a preparation method widely used for cellular cryo-electron tomography. Due to the limited ablation rates of low Ga ion beam currents required to maintain the structural integrity of vitreous specimens, common preparation protocols are time-consuming and labor intensive. The improved stability of new-generation cryo-FIB instruments now enables automated operations. Here, we present an open-source software tool, SerialFIB, for creating automated and customizable cryo-FIB preparation protocols. The software encompasses a graphical user interface for easy execution of routine lamellae preparations, a scripting module compatible with available Python packages, and interfaces with three-dimensional correlative light and electron microscopy (CLEM) tools. SerialFIB enables the streamlining of advanced cryo-FIB protocols such as multi-modal imaging, CLEM-guided lamella preparation and in situ lamella lift-out procedures. Our software therefore provides a foundation for further development of advanced cryogenic imaging and sample preparation protocols.
A: 12941.567 Å / B: 12510.182 Å / C: 6740.4 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
13.480798958333
13.480799568966
13.4808
M x/y/z
960
928
500
origin x/y/z
0.000
0.000
0.000
length x/y/z
12941.567
12510.182
6740.400
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
16
-16
250
NC/NR/NS
960
928
500
D min/max/mean
-37.881
31.106
0.485
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添付データ
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試料の構成要素
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全体 : HeLa
全体
名称: HeLa
要素
細胞: HeLa
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超分子 #1: HeLa
超分子
名称: HeLa / タイプ: cell / ID: 1 / 親要素: 0
由来(天然)
生物種: Homo sapiens (ヒト) / 株: HeLa
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
電子線トモグラフィー法
試料の集合状態
cell
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試料調製
緩衝液
pH: 7
凍結
凍結剤: ETHANE
切片作成
集束イオンビーム - 装置: OTHER / 集束イオンビーム - イオン: OTHER / 集束イオンビーム - 電圧: 30 kV / 集束イオンビーム - 電流: 0.05 nA / 集束イオンビーム - 時間: 150 sec. / 集束イオンビーム - 温度: 88 K / 集束イオンビーム - Initial thickness: 1000 nm / 集束イオンビーム - 最終 厚さ: 200 nm 集束イオンビーム - 詳細: The value given for _emd_sectioning_focused_ion_beam.instrument is Thermo Fisher Aquilos. This is not in a list of allowed values {'OTHER', 'DB235'} so OTHER is ...集束イオンビーム - 詳細: The value given for _emd_sectioning_focused_ion_beam.instrument is Thermo Fisher Aquilos. This is not in a list of allowed values {'OTHER', 'DB235'} so OTHER is written into the XML file.
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
特殊光学系
位相板: VOLTA PHASE PLATE / エネルギーフィルター - 名称: GIF Quantum LS / エネルギーフィルター - スリット幅: 20 eV