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Team:Wageningen UR/MethodsDetection
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= Detcection of VLPs = | = Detcection of VLPs = | ||
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| + | == Electron Microscopy (EM) == | ||
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| + | One of the direct methods to detect our VLPs is with Electron Microscopy or EM. We received personally a course about the EM from Jan van Lent. In this course he explained how the EM works, how you prepare the samples and how to operate the EM. Furthermore we prepared and viewed multiple samples with our newly found skill set. This all was done in the Virology department of the Wageningen UR. | ||
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| + | Electron microscopy works similar as light microscopy, but instead of visible light being used to illuminate the sample, electrons are being used. The same as light microscopy, electron microscopy uses multiple lenses to focus the beam so that the sample is properly lighted. The only differences with lenses are that the lenses of electron microscopy are electromagnetic instead of glass. EM has also a much greater resolution than conventional light microscopy. It is theoretically possible of around 0.005 nm, but in practice it is around 1-2 nm, this is because the lenses has certain errors, the operator (us) is inexperienced and the sample must be as this as possible. All these factors reduces the resolution of the electron microscope. | ||
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| + | Preparation of the sample is critical to have a good resolution, during the course we learned how to do it properly. The sample must be thin, between 2 - 300nm, and must be stable in the electron microscope, this can be done by drying or cryo-freezing the sample. After drying or freezing we stain the sample with a coating. The places where there is no coating (VLPs), electron wave can go through unhindered, while the places where there are coating, electron waves break apart. This result in a picture where you can see the particles. | ||
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| + | We checked various samples in the electron microscope. The wild types of CCMV and HepB were detected. Multiple variations of CCMV were also tested with mixed results. | ||
Revision as of 10:09, 10 September 2012
Detcection of VLPs
Electron Microscopy (EM)
One of the direct methods to detect our VLPs is with Electron Microscopy or EM. We received personally a course about the EM from Jan van Lent. In this course he explained how the EM works, how you prepare the samples and how to operate the EM. Furthermore we prepared and viewed multiple samples with our newly found skill set. This all was done in the Virology department of the Wageningen UR.
Electron microscopy works similar as light microscopy, but instead of visible light being used to illuminate the sample, electrons are being used. The same as light microscopy, electron microscopy uses multiple lenses to focus the beam so that the sample is properly lighted. The only differences with lenses are that the lenses of electron microscopy are electromagnetic instead of glass. EM has also a much greater resolution than conventional light microscopy. It is theoretically possible of around 0.005 nm, but in practice it is around 1-2 nm, this is because the lenses has certain errors, the operator (us) is inexperienced and the sample must be as this as possible. All these factors reduces the resolution of the electron microscope.
Preparation of the sample is critical to have a good resolution, during the course we learned how to do it properly. The sample must be thin, between 2 - 300nm, and must be stable in the electron microscope, this can be done by drying or cryo-freezing the sample. After drying or freezing we stain the sample with a coating. The places where there is no coating (VLPs), electron wave can go through unhindered, while the places where there are coating, electron waves break apart. This result in a picture where you can see the particles.
We checked various samples in the electron microscope. The wild types of CCMV and HepB were detected. Multiple variations of CCMV were also tested with mixed results.
