![Volumen-EM mit Rasterelektronenmikroskopie mit fokussiertem Ionenstrahl Volumen-EM mit Rasterelektronenmikroskopie mit fokussiertem Ionenstrahl]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original.image_file.100.56.file/fib-sem.jpg","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original.image_file.360.203.file/fib-sem.jpg","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original.image_file.768.432.file/fib-sem.jpg","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original.image_file.1024.576.file/fib-sem.jpg","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original.image_file.1280.720.file/fib-sem.jpg","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original.image_file.1440.810.file/fib-sem.jpg","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem.jpg/_jcr_content/renditions/original./fib-sem.jpg"})
Rasterelektronenmikroskopie mit fokussiertem Ionenstrahl
Hochaufgelöste isotrope Volumendaten für präzise 3D‑Rekonstruktionen
Schematische Darstellung eines typischen Workflows
![FIB-SEM – Abtrag FIB-SEM – Abtrag]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.100.100.0,1,1280,1281.file/fib-sem-milling.png","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.360.360.0,1,1280,1281.file/fib-sem-milling.png","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.768.768.0,1,1280,1281.file/fib-sem-milling.png","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.1024.1024.0,1,1280,1281.file/fib-sem-milling.png","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/fib-sem-milling.png","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/fib-sem-milling.png","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-milling.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/fib-sem-milling.png"})
1
Mit einem fokussierten Ionenstrahl wird so lange Material von einer in Harz eingebetteten Probe abgetragen, bis die relevante Struktur sichtbar wird.
![FIB-SEM – Bildaufnahme FIB-SEM – Bildaufnahme]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.100.100.0,1,1280,1281.file/fib-sem-image-acquisition.png","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.360.360.0,1,1280,1281.file/fib-sem-image-acquisition.png","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.768.768.0,1,1280,1281.file/fib-sem-image-acquisition.png","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1024.1024.0,1,1280,1281.file/fib-sem-image-acquisition.png","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/fib-sem-image-acquisition.png","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/fib-sem-image-acquisition.png","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/fib-sem-image-acquisition.png"})
2
Die soeben freigelegte Probenoberfläche der relevanten Struktur wird abgebildet. Materialabtrag und anschließende Bildgebung werden so lange wiederholt, bis die gesamte relevante Struktur abgebildet wurde.
![Verarbeitung der Segmentierung Verarbeitung der Segmentierung]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original.image_file.100.100.file/volume-em-processing-segmentation.png","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original.image_file.360.360.file/volume-em-processing-segmentation.png","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original.image_file.768.768.file/volume-em-processing-segmentation.png","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original.image_file.1024.1024.file/volume-em-processing-segmentation.png","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original./volume-em-processing-segmentation.png","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original./volume-em-processing-segmentation.png","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-processing-segmentation.png/_jcr_content/renditions/original./volume-em-processing-segmentation.png"})
3
Die aufgenommenen EM‑Bilder werden verarbeitet und digital zu einem 3D‑Datensatz zusammengeführt. Die Zellkompartimente lassen sich identifizieren und segmentieren.
![3D‑Visualisierungsanalyse 3D‑Visualisierungsanalyse]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original.image_file.100.100.file/volume-em-3d-visualization-analysis.png","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original.image_file.360.360.file/volume-em-3d-visualization-analysis.png","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original.image_file.768.768.file/volume-em-3d-visualization-analysis.png","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original.image_file.1024.1024.file/volume-em-3d-visualization-analysis.png","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original./volume-em-3d-visualization-analysis.png","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original./volume-em-3d-visualization-analysis.png","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/array-tomography/volume-em-3d-visualization-analysis.png/_jcr_content/renditions/original./volume-em-3d-visualization-analysis.png"})
4
Der segmentierte 3D‑Datensatz kann visualisiert, untersucht und statistisch analysiert werden.
Anwendungsbeispiele
Hochaufgelöste isotrope Visualisierung der zellulären Ultrastruktur in 3D
3D‑Imaging von HeLa-Zellen
Automatisiertes 3D‑Serien-Imaging mit der FIB-SEM-Technologie von ZEISS
Mit dem fokussierten Ionenstrahl wurde sequenziell Material mit einer Dicke von 8 nm von der Probe abgetragen – das freigelegte Blockface wurde mit einem SEM gescannt. So entstand ein hochaufgelöstes 3D‑Volumenbild. Die automatisierte Segmentierung und Visualisierung der Zellkomponenten wurde mit einem – in arivis Cloud trainierten – Deep-Learning-Modell in arivis Pro durchgeführt, sodass die verschiedenen Zellkomponenten visualisiert und quantifiziert werden konnten.
![3D‑Rekonstruktion des Golgi-Apparats einer Alge auf der Grundlage von Rohdaten nach dem FIB-Abtrag 3D‑Rekonstruktion des Golgi-Apparats einer Alge auf der Grundlage von Rohdaten nach dem FIB-Abtrag]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.100.75.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.360.270.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.768.576.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.1024.768.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.1280.960.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.1280.960.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/fib-sem/fib-sem_golgi-body_crossbeam_1.jpg/_jcr_content/renditions/original.image_file.1280.960.0,160,1280,1120.file/fib-sem_golgi-body_crossbeam_1.jpg"})
Bild mit freundlicher Genehmigung von Dr. Louise Hughes, Oxford Brookes University, Vereinigtes Königreich.
Charakterisierung des Golgi-Apparats
Erkenntnisse zur Bedeutung für Proteinmodifikation und ‑transport
Dieses Bild zeigt die 3D‑Rekonstruktion des Golgi-Apparats einer Alge aus einem FIB-SEM-Datensatz. Der Datensatz unterscheidet zwischen der cis- und der trans-Seite des Golgi-Apparats (gelb/rot: cis-Golgi-Netzwerk, violett/blau: trans-Golgi-Netzwerk). Durch die Segmentierung der Zellkomponenten aus den hochaufgelösten Datensätzen, die mit der ZEISS Crossbeam FIB-SEM-Technologie aufgenommen wurden, lassen sich die internen Komponenten präzise charakterisieren und quantifizieren.