Difference between revisions of "Serial block-face scanning electron microscopy (SBFSEM)/es"

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Las imágenes que ve en EyeWire provienen de nuestros colaboradores Kevin Briggman, Moritz Helmstaedter y Winfried Denk en el Instituto Max Planck en Alemania, y el conjunto de datos se llama [[e2198]].
 
Las imágenes que ve en EyeWire provienen de nuestros colaboradores Kevin Briggman, Moritz Helmstaedter y Winfried Denk en el Instituto Max Planck en Alemania, y el conjunto de datos se llama [[e2198]].
  
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Before any imaging could be done, the sample was stained with heavy metals. When the scanning electron microscope's electrons collided with the heavy metals in the sample, they would bounce off and they would be collected by a detector. These electrons that bounce off the sample are known as backscattered electrons. After being placed inside the chamber of the microscope, the surface of the sample is imaged. Because the Scanning Electron Microscope uses a tightly focused beam of electrons, the sample needs to be scanned in a certain pattern. The Scanning Electron Microscope will move across a line, scanning it one piece at a time, and then it will move onto the next line. This is known as raster-scanning.
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Antes de que se pudiera realizar cualquier imagen, la muestra se tiñó con metales pesados. Cuando los electrones del microscopio electrónico de barrido chocaban con los metales pesados ​​en la muestra, rebotaban y eran recogidos por un detector. Estos electrones que rebotan en la muestra se conocen como electrones retrodispersados. Después de colocarse dentro de la cámara del microscopio, se toma una imagen de la superficie de la muestra. Debido a que el microscopio electrónico de barrido utiliza un haz de electrones muy concentrado, la muestra debe escanearse en un determinado patrón. El microscopio electrónico de barrido se moverá a través de una línea, escaneando una pieza a la vez, y luego se moverá a la siguiente línea. Esto se conoce como raster-scan.
  
 
After the entire surface of the sample has been imaged, an [[Ultramicrotome| ultramicrotome]] slices off the surface of the sample and the underlying surface is then imaged in the same way. The images from the scans of each successive layer of the sample were combined to form a 3D dataset.
 
After the entire surface of the sample has been imaged, an [[Ultramicrotome| ultramicrotome]] slices off the surface of the sample and the underlying surface is then imaged in the same way. The images from the scans of each successive layer of the sample were combined to form a 3D dataset.

Revision as of 23:11, 19 July 2019

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Serial block-face scanning electron microscopy (SBFSEM) is a method for generating high resolution 3D images from biological samples. It was created at the Max Planck Institute in Germany, specifically for the purpose of imaging neurons.[1]

Cómo utiliza EyeWire SBFSEM

Las imágenes que ve en EyeWire provienen de nuestros colaboradores Kevin Briggman, Moritz Helmstaedter y Winfried Denk en el Instituto Max Planck en Alemania, y el conjunto de datos se llama e2198.

Antes de que se pudiera realizar cualquier imagen, la muestra se tiñó con metales pesados. Cuando los electrones del microscopio electrónico de barrido chocaban con los metales pesados ​​en la muestra, rebotaban y eran recogidos por un detector. Estos electrones que rebotan en la muestra se conocen como electrones retrodispersados. Después de colocarse dentro de la cámara del microscopio, se toma una imagen de la superficie de la muestra. Debido a que el microscopio electrónico de barrido utiliza un haz de electrones muy concentrado, la muestra debe escanearse en un determinado patrón. El microscopio electrónico de barrido se moverá a través de una línea, escaneando una pieza a la vez, y luego se moverá a la siguiente línea. Esto se conoce como raster-scan.

After the entire surface of the sample has been imaged, an ultramicrotome slices off the surface of the sample and the underlying surface is then imaged in the same way. The images from the scans of each successive layer of the sample were combined to form a 3D dataset.

Think of the the stack of 2D images like a flip book.The way flip books work is that from one page to another there is a small change in the drawing, and several small changes in a row create the action in the flip book. The 2D image you see in EyeWire is like one single page from a flip book. The 2D is static, but when you scroll through the slices you can see the change from one slice of retina (or page of a flip book) to another. The idea is that if you follow the shape of one neuron from one 2D slice to the next, coloring each piece as you go, you can eventually discern the shape of the neuron in the 3D. Each piece you color in the 2D builds upon the previous one. It's like stacking blocks, each layer of blocks is flat, but as you continue stacking the blocks on top of each other you eventually get a 3D shape.

For a less technical explanation you can view the article on the blog.

References

  1. ↑ Denk W, Horstmann H (2004) Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure. PLoS Biol 2(11): e329. doi:10.1371/journal.pbio.0020329
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