Jade

Bio

Hey. My name is Jade Passey, I have just finished my second year studying Veterinary Bioscience. I am now heading to North Carolina to do my placement year at the NC State Vet School.
23
Sep
2011

More Western blots this week, and still no success!!! All of this lack of success has highlighted how much work and patience goes into every aspect of a given research project.  Even though the Westerns have not woirk, the work is still incredibly interesting; it’s just a shame that recently there has been such a lack of success!! I was talking to one of the people in my lab today and she said that each Western blot costs about $200 which is a huge amount considering how many I have done!!!  It took someone in my lab three months to perfect a Tryptase Western which I repeated a few weeks ago.

In addition to the blots, which I have done for most of the week, I have been down to the necropsy room.  The necropsy room is where all the animal post mortems are done.  Watching these was fascinating, especially as I want to be a veterinary pathologist.

Early this week I also finished my cell counts- finally! I can start analysing the data next week.

Outside the lab:

This week has been pretty eventful! There was an earthquake in Virginia, which is 3 hours from here, but we felt it in Raleigh. I was sitting in the microscope room when I felt it, and it was a very odd sensation, I thought I was going insane. Hurricane Irene also hit North Carolina on Friday evening/Saturday, we are about two and a half hours from the coast which was hit hard, we just got lots of rain and winds heavy enough for blow over a few trees. It wasn’t as bad as it was forecast to be, we had a thunderstorm a few nights after the hurricane which bought heavier rain and threats of tornados.

On the Thursday before all this, we went to another Durham Bulls game, unfortunately they lost…again!!! The only two games they lost all season, and Bella and I saw them both, we figured we must be bad luck!! The second time we went, we even wore our Durham Bulls t-shirts! Again it was $1 concessions night, so we filled up on hotdogs and chips J

On Friday night we went to the Raleigh Little Theatre to see a musical called Ruthless. It was pretty weird, but still good. The theatre is tiny but the show was good. It is a spoof of musicals in general with songs and acting that are deliberately bad. We had a great evening!

On the Sunday after the hurricane the weather was amazing, so we went and explored Cameron Village some more, just to find that all the shops are boutiques and incredibly expensive, so that was a brief trip. However, we did have lunch at the Mexican restaurant again, which was delicious.

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12
Sep
2011

This week has again been much of the same work. We continue to attempt to perfect the Western Blot, which is proving increasingly difficult as the variables that we can change are now becoming limited. We have been getting results, however these are few and far between and do not appear to be reproducible, so more work needs to be done. Even the positive control is proving to be difficult to obtain results from.

On Wednesday and Friday I continued with my cell counts which are getting there and should be finished in the next week which will be good, then we have to analyse that data, which will be tricky as there will be over 700 bits of data to organise.

On Thursday I carried out another protein assay in preparation for more Western Blots next week which was pretty straight forward as I’ve done a few in the lab before, however when you have to do the assay on 40 samples, it does become a little complicated. Also on Thursday I went downstairs to learn about the work that another lab does with Zebra fish. One of the people I work with in the lab wants to do future work with these fish because they have a similar GI tract anatomy as humans in that the GI tract is in different sections. Zebra fish are also transparent which makes it possible to use fluorescent markers to mark the tract, in addition, they have rapid growth and are easy to genetically modify in large numbers. Aside from the GI work that can be carried out on these fish, Zebra fish also are one of the few animals with heart cells that can regenerate, and therefore they are becoming essential in research to cure heart disease.

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12
Sep
2011

This week has been much of the same work as last week.  However, we needed to perfect the ChAT protein Western Blot, which was previously unsuccessful.  There are a number of variables that we can change in order to optimise the assay, for example the concentrations of the antibodies. Monday and Tuesday were spent performing another Western Blot; I have been left alone to do them, with someone just around the corner in case I need help!!!

In addition to the Western Blots, I have also helped with some mouse post-mortems.  Tissues from the post-mortems were then tested using RT-PCR. The RT PCR uses mRNA to produce cDNA and then DNA with the use of primers.  The RT-PCR is used to indicate which genes are expressed under different conditions. If ordinary PCR were to be used, it would just produce DNA which would indicate the presence of genes in the genome and not their expression.

Other than that, I have been continuing with the mast cell counts, which are nearly finished, finally!

On Friday I went with someone from my lab to watch a PhD defence, which was incredibly interesting, however totally mind boggling as I didn’t understand much of what was going on! It was a great insight into how much work goes into obtaining a PhD and just how knowledgeable you have to be.

Social Activities:

It has been a pretty quiet week as far as going out goes; we didn’t do a great deal during the week and at the weekend, the weather was pretty bad. However we did go downtown to the Science Museum, which was all about the history of science in North Carolina which was fascinating. The exhibits were great and the attention to detail was unlike anything you would see in the UK. Afterwards we went for some lunch in a little cafe and then my housemate Kristen picked us up and gave as I tour of downtown Raleigh in the car. In the afternoon we went to the cinema on campus with a few people from my lab to see ‘Crazy Stupid Love’ which doesn’t come out in the UK for a few months… amazing film!  We then found a frozen yoghurt shop, which is the healthy alternative to ice cream, however they have massive pots, which of course have to be filled  and a whole selection of unhealthy toppings which we piled on!! On Sunday we went back to the mall to go to the Cheesecake Factory again, this time we decided we would not eat a big lunch so there would be room for cheesecake, so we order a starter of nachos which were massive, and then we got cheesecake!

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24
Aug
2011

This week I have repeated another protein assay with different samples collected from pig intestines. The assay has shown that the optimum dilution of the samples is 1:100, which also fits perfectly into the standard curves that have been created. Additional Western Blots will be performed over the next few weeks. Continue reading »

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10
Aug
2011

This week has been very busy in the lab. I have been able to use some of the skills that I have learnt in the Biochemistry practicals at University of Surrey, so this was really good. The beginning and the end of the week were spent cutting tissues and staining the tissues in order to perform mast cell counts. I have made about 150 slides already! Continue reading »

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26
Jul
2011

We are having a fantastic time, the weather is incredible, and we are currently having a heat wave so temperatures are exceeding 100oF!

The social life is good and the lab work is incredibly interesting. The day after we arrived we went straight to the University and met the scientists we will be working with for the next 3 months and were guided through the type of work we would be undertaking. I am working in the Gastrointestinal Physiology department where they are researching the effects of stress as a result of early weaning of piglets. In these studies the effects of the stress are determined by the number of mast cells present in tissues and the nature of the Mast cell degranulation.

On my first day I was taught how to cut tissue that had previously been treated and frozen into sections, so by the second day I was cutting the tissues on my own and mounting them onto slides prior to staining. I also learnt the protocol for immunofluorescence, which is fairly complex and time consuming, but when the slides are viewed under a fluorescence microscope the images are incredible. For the rest of the week I have cut tissues on the Cryostat in order to create slides which I have then stained.  The stain colours the majority of the tissue blue, however it stains the mast cells purple, which enables them to be easily counted under the microscope. In the afternoons I have been counting these mast cells and imputing the data into a spread sheet in order for it to be analysed at a later date.

The people I am working with in the labs are so helpful and friendly and so willing to answer any questions I have. We have been out for lunch a couple of times, which has helped me get to know everyone better.

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