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<img src="https://static.igem.org/mediawiki/2018/e/e6/T--Rheda_Bielefeld--StickstoffFichte400.jpg" style="width:400px;height:auto;"></img><br> <br> pollen of a spruce opened with nitrogen with 400x magnification (light microscope) | <img src="https://static.igem.org/mediawiki/2018/e/e6/T--Rheda_Bielefeld--StickstoffFichte400.jpg" style="width:400px;height:auto;"></img><br> <br> pollen of a spruce opened with nitrogen with 400x magnification (light microscope) | ||
</article> | </article> | ||
+ | <h2> Electron Microscopy </h2> | ||
+ | <article> | ||
+ | There are two different kinds of microscopes: <br> | ||
+ | <ol> | ||
+ | <li> Light Microscope </li> | ||
+ | <li> Electron Microscope </li> | ||
+ | </ol> <br> | ||
+ | While the light microscope is a tool used in basic biology lessons in school to see plant cells and seeds, the power of a light microscope was not enough to check whether the pollen were cracked or not because the pollen cores are too small. To see better results, we needed a electron microscope. | ||
+ | <br> <br> | ||
+ | An electron microscope uses a beam of electrons to create an image of objects. ELectrons are thrown on an object and are reflected to a detector. The given data are be analyzed to form a picture. <br> | ||
+ | That would have helped us to see the success of our experiments- if it had worked so far. | ||
</div> | </div> | ||
Revision as of 09:02, 17 October 2018
Extracting Pollen
Opening Pollen
To form a trypsin solution, we resuspended 20µg of trypsin in 200µl of H²0. Then we split up the solution into 4 eppis with 50µl each, having approximately 5µg of trypsin within every eppi. According to our proportionality-calculations we then added pollen to the trypsin and incubated at 37°C overnight. The following day we analysed our samples with light microscopy, enlarging by 100 and 400.
To use ribolyser tubes we added 10, 20 and 50 mg of pollen combined with 400 µl of water into the tubes. In the next step we shook the tubes in the ribolyser and centrifugated at 110 rpm for 10 minutes. The results were two different layers, one brighter than the other. On the top layer was white shroud. After abstracting the light-brown, lower, layer with a chemical dropper, 400µl lysebuffer were added and the solution was centrifugated for 1 minute with 11000 rpm. Afterwards, we used the machery nagel plant Nucleo spin III Kit for the DNA extraction.
Lastly, we tried to open the pollen with liquid nitrogen. Therefore, we put pollen in a small bowl and added liquid nitrogen. Then we cracked the pollen using mortar and pestle and checked the results of all approaches with the light microscopy.
pollen of a spruce opened with nitrogen with 400x magnification (light microscope)
Electron Microscopy
- Light Microscope
- Electron Microscope
While the light microscope is a tool used in basic biology lessons in school to see plant cells and seeds, the power of a light microscope was not enough to check whether the pollen were cracked or not because the pollen cores are too small. To see better results, we needed a electron microscope.
An electron microscope uses a beam of electrons to create an image of objects. ELectrons are thrown on an object and are reflected to a detector. The given data are be analyzed to form a picture.
That would have helped us to see the success of our experiments- if it had worked so far.
Gallery
(light microscope) pollen of a spruce with 400x magnification
(light microscope) pollen of a birch with 100x magnification
(light microscope)