Honors Experience Journals

CCHMC Neurology Research

Cassie Bivens

February 20, 2018

As of today, I have been working in the Skelton lab at Cincinnati Children’s
Hospital for two weeks. It has definitely been a new experience for me so far, even
though I worked in a neighboring lab with a different postdoc for a year and a half.
My postdoc for this experience is Dr. Marla Sunderman, who has asked me to call
her Marla. I have been coordinating with her as well as Zuhair Abdulla, a
neuroscience graduate student. Marla’s study looks at how genes play a role in
Bipolar Disorder. The part of the study that I have been assigned to is doing Western
blots, which use fluorescent antibodies to show whether or not certain proteins are
present in samples of certain rat brain cells.
So far, I have successfully completed one Western blot using cells prepared
from culture. These are B35 cells, which come from neuroblastomas in rats. A
Western blot is a consecutive two-day process. When the cells are ready to be
harvested, I harvest them from culture using a protocol Marla gave me. Then, I run a
protein assay, which tells me how much protein is in various dilutions of the cells.
This allows us to find the right dilution to use for the blot that will maximize the
results. Then, I load each sample in its proper dilution into a specially designated
gel. This is hooked up to an electrophoresis machine and allowed to run for 35
minutes. After 35 minutes, the bands of sample have run down to the bottom of the
gel. The gel is then removed and goes through a two-hour process of transferring
the bands from the gel to a special membrane. After the transfer step has occurred,
the membrane is soaked in a blocking buffer for one hour. After that, primary
antibody solution is added to the membrane, which then sits on a rocker in a cold
room overnight.
The next day, I come back in to the lab and go through various timed rinsing
steps with the membrane. I then add the secondary antibody before using a special
machine to read the results of the blot. The results come in the form of fluorescent
bands that appear on the image obtained by the machine. They tell us whether
certain proteins are present. So far, we have found phospho-DRP and actin to be
present in the cells.
Working closely with the graduate students in the lab has given me some
insight into what it would be like to pursue a graduate degree in neuroscience
research, which is something that I have previously considered. I will definitely be
thinking about my options throughout the course of the semester as I continue to
see what sorts of things graduate students do.

March 6, 2018
I have done several Western blots in the past two weeks, each with better
results than the last. The more I do Western blots, the more I understand how I can
maximize the results in the future. Since it is such a long process and there are so
many tiny steps that can go wrong along the way, it is really important to keep
diligent notes about EVERYTHING. I am learning that this is a good practice for
nearly any laboratory procedure. That way, if something doesn’t turn out as
expected, you can go back and see what you may be able to do differently to obtain
better results next time. For example, by keeping good notes and tweaking my
methods I was able to significantly reduce “background noise” in my Western blots
and make the images much clearer overall. This is definitely a very important part of
the scientific process; it is vital to be able to make mistakes and learn from them.

March 20, 2018

I have been coming in to the lab several days a week to work on Westerns,
and I love that I can see my technique improving! Each time I do a blot, I get more
efficient and am able to tweak little details to get better results. For example, the last
couple of blots have had issues with air bubbles, and by introducing a new step I
was able to minimize the air bubbles to get a clearer reading. I love that I am able to
take pride in my work and contribute to the scientific community in this way, and
that the results I am getting will end up being useful to the study itself.
All in all, I am focusing on bettering my techniques, especially since Western
blots are a very common method used in research. I am hoping that having this skill
will come in handy in any future research endeavors. Zuhair has been really helpful
with identifying things that could be done better, since he has done so many
Western blots in the past and has run into just about every possible problem that
could be had. I am definitely learning just how specific nearly every scientific
process is and how careful you really have to be at all times. It is really amazing how
the smallest details can have such a big impact!

April 3, 2018
For the last two weeks, I have continued to do Western blots. However, some
of them have not been turning out very well with the antibodies we have been using,
so we will be switching from phospho-DRP and actin to phospho-DRP and Reelin as
the primary antibodies. For some reason, the actin band has been appearing, but it
has been in the wrong location on the gel, where we would not expect an actin band
to be. Marla will be contacting the company that makes the antibodies, because it is
likely that we were given a defective batch of antibody. Usually when this happens,
the company sends a free batch, which is important because each bottle of antibody
contains only 100 microliters and costs hundreds of dollars. For reference, 100
microliters is just 0.1 milliliters—a tiny amount! It is really amazing to me how
massive of a budget each laboratory must have in order to maintain their research.

April 17, 2018

As my time in the lab for this semester is coming to an end, I am beginning to
more strongly consider pursuing a graduate degree in neuroscience. I have always
been very interested in the human brain, but I never expected to become so
fascinated with research. I always just assumed that the only career you could have
that involved the human brain was to become a neurological physician. However, I
am really glad that my eyes have been opened to see that maybe a career in
neuroscience would be both a viable career path and a way to work in a field that I
have always been passionate about. It is definitely worth considering.
 This semester has really taught me a lot about improving upon scientific
skills. It’s easy to say that you know how to do something because you’ve done it
before, but it’s more difficult to critique yourself and see that just because you know
how to do something, doesn’t mean that there aren’t things you can improve upon.
This applies to many things in life, in addition to scientific research. The ability to be
self-critical is a very important life skill that I feel I have really been able to develop
this semester.

May 1, 2018
For my last journal entry, I would like to reflect on just how far I have come
in a single semester. At the end of last semester, I was being taught how to perform
Western blots, and I was supervised every step of the way. This semester, I have not
only successfully performed Western blots completely independently, but I have
been able to improve upon my results. Being able to take pride in my work is such a
great feeling, and it shows me that there is always something more that I can learn. I
have also gained a sense of self-confidence that I did not have before, because I have
been entrusted to perform important experiments completely independently. It
means a lot that Marla trusts my judgment and skill enough to put me in charge of
Western blots for her study.
Marla has told me that I am always welcome in the lab, whether it is this
summer or in the fall when school is back in session. I am really grateful for the
opportunity that I have had to work with her this semester, and I would absolutely
like to work with her in the future! She is a great mentor and an impressive scientist,
and her work has inspired me to consider neuroscience research as a viable career
path.