![EM de volumen con FIB-SEM criogénica EM de volumen con FIB-SEM criogénica]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original.image_file.100.56.file/cryo-fib-sem.jpg","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original.image_file.360.203.file/cryo-fib-sem.jpg","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original.image_file.768.432.file/cryo-fib-sem.jpg","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original.image_file.1024.576.file/cryo-fib-sem.jpg","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original.image_file.1280.720.file/cryo-fib-sem.jpg","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original.image_file.1440.810.file/cryo-fib-sem.jpg","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem.jpg/_jcr_content/renditions/original./cryo-fib-sem.jpg"})
FIB-SEM criogénica
Capte una instantánea de la dinámica celular en estados casi naturales con ultrarresolución
Representación esquemática de un flujo de trabajo típico
![Fresado de FIB-SEM criogénica Fresado de FIB-SEM criogénica]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.100.100.0,1,1280,1281.file/cryo-fib-sem-milling.png","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.360.360.0,1,1280,1281.file/cryo-fib-sem-milling.png","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.768.768.0,1,1280,1281.file/cryo-fib-sem-milling.png","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.1024.1024.0,1,1280,1281.file/cryo-fib-sem-milling.png","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/cryo-fib-sem-milling.png","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/cryo-fib-sem-milling.png","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-milling.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/cryo-fib-sem-milling.png"})
1
Con un FIB se fresa una muestra vitrificada y sin tinción hasta que la estructura en cuestión se hace visible.
![Adquisición de imágenes de FIB-SEM criogénica Adquisición de imágenes de FIB-SEM criogénica]({"xsmall":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.100.100.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png","small":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.360.360.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png","medium":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.768.768.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png","large":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1024.1024.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png","xlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png","xxlarge":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png","max":"https://www.zeiss.com/content/dam/rms/reference-master/applications/life-sciences/volume-em/cryo-fib-sem/cryo-fib-sem-image-acquisition.png/_jcr_content/renditions/original.image_file.1280.1280.0,1,1280,1281.file/cryo-fib-sem-image-acquisition.png"})
2
Se capturan imágenes de la superficie de la muestra recién expuesta de la estructura de interés. Este proceso de fresado y captura de imágenes se repite hasta que se ha capturado una imagen completa de la estructura. La muestra permanece vitrificada durante todo el proceso.
![Segmentación de procesamiento Segmentación de procesamiento]({"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
Las imágenes de microscopio electrónico adquiridas se procesan y alinean digitalmente en un conjunto de datos en 3D. Los compartimentos celulares se pueden identificar y segmentar.
![Análisis de visualización en 3D Análisis de visualización en 3D]({"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
El conjunto segmentado de datos en 3D se puede visualizar, estudiar y analizar estadísticamente.
Ejemplo de aplicación
Comprender el proceso de biomineralización de cristales de calcita en un cocolitóforo
Formación de cocolitos en el cocolitóforo Emiliania Huxleyi
Visualización de partículas de calcita y su entorno ultraestructural
Es difícil capturar imágenes del entorno ultraestructural de las fases cálcicas solubles y amorfas de los cocolitóforos con los protocolos clásicos de preparación de base acuosa. En condiciones criogénicas, el FIB-SEM permite captar imágenes de algas marinas vitrificadas en estado casi natural, dilucidando la ultraestructura en 3D.
Las células de E. huxleyi se congelaron a alta presión. Durante el proceso de captura de imágenes, la muestra se mantuvo en estado de congelación; el conjunto de datos FIB-SEM se adquirió utilizando un ZEISS Crossbeam en condiciones criogénicas. La reconstrucción en 3D muestra los cocolitos maduros (amarillo), un cocolito in statu nascendi (azul) y cuerpos lipídicos (rojo).