Enhanced FIB-SEM systems for large-volume 3D imaging.
C Shan XuKenneth J HayworthZhiyuan LuPatricia GrobAhmed M HassanJosé G García-CerdánKrishna K NiyogiEva NogalesRichard J WeinbergHarald F HessPublished in: eLife (2017)
Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) can automatically generate 3D images with superior z-axis resolution, yielding data that needs minimal image registration and related post-processing. Obstacles blocking wider adoption of FIB-SEM include slow imaging speed and lack of long-term system stability, which caps the maximum possible acquisition volume. Here, we present techniques that accelerate image acquisition while greatly improving FIB-SEM reliability, allowing the system to operate for months and generating continuously imaged volumes > 106 µm3. These volumes are large enough for connectomics, where the excellent z resolution can help in tracing of small neuronal processes and accelerate the tedious and time-consuming human proofreading effort. Even higher resolution can be achieved on smaller volumes. We present example data sets from mammalian neural tissue, Drosophila brain, and Chlamydomonas reinhardtii to illustrate the power of this novel high-resolution technique to address questions in both connectomics and cell biology.
Keyphrases
- high resolution
- electron microscopy
- liver fibrosis
- electronic health record
- deep learning
- single molecule
- endothelial cells
- mass spectrometry
- big data
- single cell
- convolutional neural network
- optical coherence tomography
- white matter
- high speed
- cell therapy
- tandem mass spectrometry
- cerebral ischemia
- brain injury
- fluorescence imaging