Summary Lab Report
In the lab week, we learned several techniques to extract DNA, inject viruses into neurons to make them visible and to make embryos better visible under the microscope.
The first day
We learned the basic techniques to grow neurons cells extracted from an E8 chicken embryo.
We worked under the laminar hood for a sterile environment. Under the hood, we prepared the neurobasal media which will later contain the neurons.
We then opened the eggs using scissors, extracted the embryo and removed all body and brain parts to extract telencephalon containing of both hemispheres.
Then we cut the hemisoheres into very small pieces to then seperate the neurons with accumax.
Centrifugation then isolated the neurons from the rest.
Then we put the neurons on a petri dish and the neurons got infected with a virus (done by the lab personnel)
Second day
We did in vivo work on the E3 chicken embryo. The goal was to localize the embryo and inject a vehicle to make the embryo visible.
First we prepared the aspirator tube with a neutral red solution. Then we opened up the eggs with creating a lid with scissors and attached it to the egg with tape.
then we located the embryo under the microscope and injected the red solution
Third day
We learned how to tag our cultured cells from day 1 with antibodies. +
Under the laminar hood, we fixated the cells by using PFA. Once they were attached to the bottom of the well, we washed the well with PBS:
Then we transferred the neurons to a petri dish and applied a blocking solution.
Then we prepared the antibody
Fourth day
We continue on the work from wednesday (day 3). We did a lot of washing this day.
We removed the auntibody from the coverslip and put the coverslip back into the well.
So we washed the coverslip with PBS.
Then we put it back on the parafilm, we got another antibodies on the coverslip and inculated it over night.
Then we did DNA extraction:
then we took the injected eggs from tuesday and opened up the eggs. Some embryos sadly died, so we only used the ones that were still alive.
Then we put the embryo in a petri dish, cut off the head and removed all of the surroundings of the brain.
Then we sliced uop the brain into small pieces. We then added Proteinase and after that centrifuged to do separte DNA from the rest.
You could then clearly see the white mass being the extracted DNA which we than tranferred into a clean tube.
Fith day
On the last day we took our coverslips from th fridge, removed the antibodies and cleaned it again with PBS.
then we dropped a small amount of DAPI on the slide.
Then we could observe the neurons under the microscope. We could zoom in 20x or 40x and see with the different filters different structures. With DAPI (UV filter), we could observe the nuclei, with GFP and a blue light filter we could see the tubulin, which showed us the enitre structure of the neurons.
Two groups even found neurospheres, a dense collection of many neurons at one location.
see also
Tags: neurobiology science
Superlink: 051 ☣Neurobiology 050 🧠Neuroscience
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Created: 2025-06-11 09:58