Neuroscience 2010
Tuesday, January 11, 2011
Shadowing
Today Ryan and I had the privilege of shadowing each other. He did not take Bio 115, but he did catch on quickly when I showed him and explained to him what I was doing. I went into his lab absolutely clueless. I knew he was doing something with biodiesel, but was not sure what he was actually doing. He is currently comparing yields of biodiesel based on what method he used to see which one is most efficient. I observed him immobilize an enzyme onto celite. He told me that this not only increases enzymatic activity, but it is also reusable during the activity. The enzyme attaches to the celite and is filtered out through vacuum filtration leaving it for Ryan to reuse again.
Ryan previously made a sodium phosphate buffer to help dissolve (He uses 1 L for this process).
Using liquid nitrogen, Ryan performs lyophilizing (a process known as flash freezing). It is mainly a dehydration process that Ryan uses in order to allow the frozen water to sublime and go from a solid to a gas. While I was there he was in the process of freezing, but I did not stay long enough to see him add heat to it, thus converting his product.
Ryan also uses a Roto-back vacuum for his enzyme. This works as a sublimation in which all water is sucked out leaving his enzyme as a dry powder (Celite, enzyme, and salt remain). The enzyme at this stage is attached to the celite.
Ryan then uses the immobilized enzyme and incubates it with 95% ethanol and soybean oil. He sets it out on a watchglass to evaporate which he said is mainly a timing issue. Then he boils out all the ethanol (alcohol boils at a lower temperature compared to oils). He spoke of the importance of using boileezers (that should bring back some organic chemistry memories). Once this has been done, Ryan can then perform vacuum filtration in which oils are sucked in. Then he boils the enzyme and watches for the temperature to change. Once the temperature rises above 100 degree Celsius, he knows it just contains oil. Ryan also explained how weird it was that if you boil it, it turns brown…but if you evaporate it, it stays a clear color. This is just one unique physical property.
After spending some time in the inorganic lab today I am not so sure I would enjoy it. I could understand what Ryan was doing, but I was very uninterested in it mainly because I was not real familiar with his study. He was patient with me as I asked him many questions and he explained every technique thoroughly which was helpful in fully grasping what he was doing. I kind of realized how “timing is everything” for his research as well. I now have a fuller understanding for what he is doing and how beneficial this type of research is to the science community.
Article Review...SLEEP ANYONE?
In the article, Why Does Lack of Sleep Affect Us Differently? Study Hints It May Be In Our Genes, discusses a study in which people who have a gene linked with narcolepsy are monitored. 92 people without this gene variant and 37 people with this gene variant did not display any type of sleep issues during the monitoring process. During the process, subjects came to the lab and slept for 10 hours in a bed and reported feeling rested. During time trials where the subjects were constantly woken or disturbed, subjects were only allowed to sleep for 4 hours a night. In order to keep the subjects from falling asleep, researchers would do anything to entertain them.
Subjects were allowed to play card games, read, watch movies, anything to distract them for how tired they were. Sleep quality was measured and subjects also rated their sleepiness, and performed memory tests.
Researchers found that those with the variant of the gene were found to much more sleep deprivation and reported feeling more tired in both conditions: being fully rested, and running on four hours of sleep. People with the gene variant were also getting up multiple times during the night/ their sleep trial as opposed to those without the gene variant.
According to the actual researcher, this gene will help identify those people who are more susceptible to falling asleep and feeling sleep deprived. The researcher felt that this information would be most beneficial to people who work late night shifts, have severe family obligations/ are taking care of children or elderly people who might waken during the night, and people who travel through different time zones for work or personal obligations.
I did not like this review of the article because I felt as if the writer jammed together the methods and results and threw it on paper as fast as possible. When I am reading I like to know everything in order. The first thing that disturbed me was the methods. The writer did not explain the technique(s) or the process in which patients recorded their sleepiness.
I did not know if they used a survey format or if the patient met individually with the researcher and spoke of their feelings. I also feel as if the activity level of the patients should have been recorded. That might be key to determine at what point people started to “slouch” or “feel too tired to function.” I did not know age or gender either, both of which are vital to a sleep study.
The article mentioned no previous training required by subjects and I wondered how the subjects prepped for something like this. Would they sleep a lot in order to feel rested going into this trial? Would they have “practiced” on their own? This article was originally from October and I remember it coming across my aol news feed and the title caught my eye, Why Does Lack of Sleep Affect Us Differently? Study Hints It May Be in Our Genes. I was intrigued by how researchers were studying sleep and if new improvements had been made to help patients or people in general get a better quality night sleep.
Sleep fascinates me and maybe it is because it is lacking in my life and so I feel the urge to read about it. It would be interesting to tie a sleep study with food or a sleep study with beverage and see how that impacts sleepiness. I am guilty of eating something just to stay awake and drinking multiple cups of tea just to help hold my eyes open.
Tuesday, January 4, 2011
Technique
One technique that I am using in my research is creating vials that shelter the Drosophila after they have been exposed to the cadmium. It takes, on average, ten days after the flies have been collected and stored in their respective concentration of cadmium to produce usable offspring that could potentially contain virgin flies. During the ten day process, it is typical to see larvae and pupae at about day four (Pupae are brown and are an older version of larvae). Once there is an abundance of flies, the parents are etherized and either dumped or used in other experiments (such as survivability). Bottles are then cleared in which all parent flies are disposed. This ensures that when bottles are observed (usually three to four hours later) virgin flies will only be in the bottles. Virgin flies are typically light in body color and can be seen with the naked eye.
The technique that I was primarily writing about was creating vials for these virgin flies. It is important to allow the flies to leave their “cadmium contained” home and be isolated from all other flies. This ensures that no mating will occur before an actual test of research observed mating occurs. Dr. Schnee and I isolate the flies not only by gender but also individually. It is a tedious process in which small glass vials are used containing .8 g of Carolina Biological Supply (Drosophila formula) and .3 mL of water. Once bottles have been prepared and labeled, virgin flies are placed individually into these vials by use of an aspirator. I currently have approximately 30 vials that have been prepared over the last two days that will be used for testing on Wednesday. The flies that were used were flies that were collected a week ago Monday while the science hall was first experiencing its heating problem (although, I did not realize it when I was in the fly room). J These flies do not need to be held in the incubator, they are fine to leave at room temperature.
Some experiments suggest four days instead of three, but I am simply testing on three day intervals due to our shared research schedule with two other science friends.
The most difficult part of this process of using virgin flies and testing their mating behavior is getting one female and one male into a vial. I typically look to if there is an abundance of prepared vials (where flies are held individually) and use those flies as my moving organisms to other vials. Losing flies in the process is common and irritating if there are a scare number of flies available. Using an aspirator I move one fly to another vial containing a fly. It is possible to move both flies to a new vial that does not contain any food and Dr. Schnee and I will probably try both methods. Once this process is complete, Dr. Schnee and I record mating behaviors and courtship time.
*Survivability (mentioned above in this blog) will be discussed in a future post.
*Mating behaviors and courtship will be discussed in a future post.
*I attached the article that I brought to class today. The intro is most beneficial to my research because it talks about the effects cadmium can have on Drosophila in terms of motor and nerve function. This could explain why certain levels of cadmium are hindering to the flies and don’t allow for successful mating to occur. The article goes into greater depth on nerve functioning and it suggests that perhaps cadmium was placed at the wrong part of the neuron when trying to determine its affect on calcium channels (which is also associated with muscle and nerve functions). Feel free to read!
*Here is a picture of a vial containing one fly taken during research time today. Enjoy!
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