Stephanie Wu | Week 3 | More Slicing and Mounting
06.19.17 - 06.25.17
For the majority of this week I spent my time slicing and mounting the brains Felicia had previously removed from some mice via perfusion after inserting the injections she was working with. The main type of mouse I have worked with are the rAAV2 Delta-Cre mice (which are the controls) and the rAAV2 GFP-Cre mice (which are the experimentals), which are then divided into hetero and homo. From there, Felicia targeted specific regions of the brain, mostly injecting into areas such as the nucleus accumbens (NAc) and the dorsal striatum (DS). She injected both a control virus as well as multiple experimental viruses. By slicing the brains we would eventually be able to image them on the microscope and see the effects of the injection on the synapses of the brains and the different afferent sites as well.
Besides slicing and mounting the brains, I also accompanied one of the post docs, Kyuhyun, downstairs where the mice are kept. There I helped him with weaning Felicia's cages, his behavior experiment, and watched him perform two surgeries.
Weaning is the act of separating mice from their parents at postnatal (PND) day 14 (so 14 days after they are born). I helped by removing the mice from the breeding cages and separating the two genders into two new cages. You can identify the gender by looking at the underside (female mice typically have two white spots on their hing legs, while male mice don't). After separating them, I helped write new cage cards for identification, including things like their date of birth and the parents' genotypes.
Afterwards I watched as Kyu conducted his behavior experiment and also watched him inject his own virus into the mice brains during a surgery, and helped with the injection process and keeping track of their vitals. In preparation for the surgery he has to put the mouse under a type of anesthetic, and in order to prevent the mouse from dying he places it on a heating pad to reduce the lowering of its body temperature. He also makes sure that the head is secure and doesn't move during the procedure as well as level it so that the injection is injected correctly. He uses coordinates to find the injection sites and some markings on the mouse's brain called bregma to find the right area. He also makes sure to put a special type of cream on the mouse's eye to prevent infection as the mice are unable to blink during the surgery. To reach the brain they typically use Nair to remove the hair, then cut it open and level it for the injection. Afterwards, they suture (stitch) the skin on the head back together, I was able to do this myself! Overall procedure for the surgery is the same, just the injection and injection sites are what differ.
For the most part, a lot of my time this week was focused on slicing and mounting the brains as the whole procedure takes a while to complete.
On the weekend I visited Temple University with Pang, as well as walked around the city. Pang and I also visited a cafe and had bingsoo, which is a Korean shaved ice dessert. This week overall was great and I'm looking forward to next week and Felicia returning from visiting her home in California!
For the majority of this week I spent my time slicing and mounting the brains Felicia had previously removed from some mice via perfusion after inserting the injections she was working with. The main type of mouse I have worked with are the rAAV2 Delta-Cre mice (which are the controls) and the rAAV2 GFP-Cre mice (which are the experimentals), which are then divided into hetero and homo. From there, Felicia targeted specific regions of the brain, mostly injecting into areas such as the nucleus accumbens (NAc) and the dorsal striatum (DS). She injected both a control virus as well as multiple experimental viruses. By slicing the brains we would eventually be able to image them on the microscope and see the effects of the injection on the synapses of the brains and the different afferent sites as well.
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| (some brain slices (N1C. 87)) |
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| (more slices) |
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| (some mounted slides of N1C. 125 & 126 brain slices) |
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| (what my slide box currently looks like) |
Besides slicing and mounting the brains, I also accompanied one of the post docs, Kyuhyun, downstairs where the mice are kept. There I helped him with weaning Felicia's cages, his behavior experiment, and watched him perform two surgeries.
 |
| (Kyuhyun working on his behavior experiment) |
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| (some of his mice) |
Weaning is the act of separating mice from their parents at postnatal (PND) day 14 (so 14 days after they are born). I helped by removing the mice from the breeding cages and separating the two genders into two new cages. You can identify the gender by looking at the underside (female mice typically have two white spots on their hing legs, while male mice don't). After separating them, I helped write new cage cards for identification, including things like their date of birth and the parents' genotypes.
 |
| (weaning the mice) |
Afterwards I watched as Kyu conducted his behavior experiment and also watched him inject his own virus into the mice brains during a surgery, and helped with the injection process and keeping track of their vitals. In preparation for the surgery he has to put the mouse under a type of anesthetic, and in order to prevent the mouse from dying he places it on a heating pad to reduce the lowering of its body temperature. He also makes sure that the head is secure and doesn't move during the procedure as well as level it so that the injection is injected correctly. He uses coordinates to find the injection sites and some markings on the mouse's brain called bregma to find the right area. He also makes sure to put a special type of cream on the mouse's eye to prevent infection as the mice are unable to blink during the surgery. To reach the brain they typically use Nair to remove the hair, then cut it open and level it for the injection. Afterwards, they suture (stitch) the skin on the head back together, I was able to do this myself! Overall procedure for the surgery is the same, just the injection and injection sites are what differ.
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| (fooling around before surgeries featuring Kyu) |
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| (one of Kyu's mice in the middle of the injection and surgery) |
For the most part, a lot of my time this week was focused on slicing and mounting the brains as the whole procedure takes a while to complete.
 |
| (putting my finished slides for the day away) |
On the weekend I visited Temple University with Pang, as well as walked around the city. Pang and I also visited a cafe and had bingsoo, which is a Korean shaved ice dessert. This week overall was great and I'm looking forward to next week and Felicia returning from visiting her home in California!
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| (image of the river near my apartment) |
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| (green tea bingsoo) |
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