Honors Journals

Fall Semester 2017

Undergraduate Research Experience
Cassandra Bivens

8/28/17
For the past two weeks, I have been back in the lab, working with Dr. Jenna
Sprowles on the study I have been working on for the past year. This study looks at
how SSRI’s (a class of antidepressants) consumed during pregnancy can affect the
offspring throughout childhood and adolescence, using the hypothesis that it will
cause changes in brain structure and behavior similar to those in Autism Spectrum
Disorder, or ASD. For the past year, I have helped Dr. Sprowles (or Jenna, as she
likes to be called) with both animal and laboratory procedures, since the study uses
a rat model to translate to how SSRI’s may also affect humans. When I first started
working in the lab, I began with dosing the rats with antidepressant through
subcutaneous injection. After gaining some experience with that, I learned how to
run behavioral tests on the rats through the use of water mazes. Now, however, the
study is in its final months, so behavior is over and dosing will soon be done. Jenna’s
contract with the hospital is up at the end of December, so she has planned her
study accordingly to be just about finished by then.
At the end of last year, I began working on more laboratory techniques, such
as cresyl violet staining, protein assays, and sectioning slices of brain tissue with a
Cryostat. These procedures are what the rest of the study will mainly focus on, so I
have been doing a lot of cresyl violet staining and sectioning in addition to dosing
within the last two weeks. I have also learned a new method of mounting the brain
slices on microscope slides that is much more challenging, so it will be my goal to
improve upon that method.

9/11/17
I have been working in the lab quite a bit lately, and I think that I have been
really getting the hang of some of the procedures! I have been working a lot on
cresyl violet staining, sectioning, and taking slide images with a microscope. I really
enjoy the cresyl violet staining, which uses a multi-step process to stain brain slices
mounted onto microscope slides a dark purple color. This makes the outlines of the
brain structures we want to look at, specifically the hippocampus (the center for
learning and memory), to appear much more defined, since the areas with the
highest protein concentration are stained a much darker purple color than the areas
with less protein. Essentially, it’s a way of visually outlining the structures of the
brain.
I have also been doing quite a bit of sectioning with the Cryostat, which uses
cryogenic freezing to solidify the rat brains so that they can be sectioned into slices
that are several thousandths of a millimeter thick. These are then mounted onto
slides, which can be done two ways: wet or dry. The wet method is much more
difficult to master, but it has its benefits for certain types of studies.
Slide imaging is the only thing I have been working on that I have not done
previously. Once the brain slices are mounted on the microscope slides and stained,
they are placed under a microscope that has a camera attached to it. As I mentioned,
our study focuses on the hippocampus, so I have to locate that within each slice.
Then, I have to take digital images of each of three regions of the hippocampus on
each side of the slice. These three regions are called the CA1, CA3, and dentate gyrus,
or DG regions. I really enjoy this procedure as well, because it allows us to compare
images of hippocampal structures from control groups with those from rats who
were dosed with the antidepressant. This allows us to see if there are any physical
differences in brain structures that correspond with SSRI consumption.
I have learned a lot in the last two weeks, and I really hope that I continue to
be able to learn more laboratory techniques as dosing winds down!

9/25/17
In the last two weeks, I have been working on cresyl violet staining,
sectioning brain tissues, slide imaging, and protein assays. I did one protein assay
with Jenna last year, and the results were not nearly what I wanted, because they
indicated that a large amount of error was present on my part. This time, however, I
was able to correct my previous mistakes, and my protein assay turned out very
well. In fact, it turned out so well that I printed out the results to hang on my fridge!
Protein assays are a lengthy process that involve taking samples of ground-up brain
tissue that are suspended in solution and making sure that each sample has the
same amount of protein in it—essentially, it’s a standardization process. This then
allows the samples to be used for Western blots, which need all samples to have the
same “weight” of proteins in them for the procedure to work properly. Jenna has
assured me that I will get to work on Western blots at some point this semester as
well.
I have been in the lab more this semester than I was last year, and with 18
credit hours it can be pretty difficult to balance sometimes, so I have definitely had
to learn to manage my time well. Sometimes it can seem like there’s just not enough
time in a day! However, I think that my time management skills have improved as a
result.

10/9/17
For the last two weeks, I have been working on cresyl violet staining and
slide imaging. Sectioning with the cryostat has been wrapped up for the rest of the
study, so now we have to stain the brain slices so that they can be imaged.
Essentially, once the brain is obtained from the rat, there are four steps that must be
taken. The brain must be sectioned into slices, which are then mounted onto
microscope slides. The slides must then go through cresyl violet staining so that
they can proceed to the final step, which is slide imaging. Without cresyl violet
staining, slide imaging would be unsuccessful, because we would not be able to
properly see the structures we are looking for. Since we have finished with all of the
sectioning and mounting that needs to be done for the study, we have done a lot of
work to catch up on staining and imaging the slides that were obtained weeks and
even months ago. These slides can remain in the freezer indefinitely, so some of
them were sectioned and mounted quite a while ago and were simply waiting for
someone to become available to perform the next steps.
 I really enjoy the feeling that comes with mastering new laboratory
techniques, so I appreciate the extra practice with slide imaging, since that is my
most recently acquired skill. I am also excited that dosing will soon be wrapping up,
because that means that I can spend my time focusing on laboratory techniques
rather than rat injections. Dosing isn’t terrible by any means; I just really enjoy
working in the lab more than I enjoy working with the animals.

10/23/17
Dosing has finally wrapped up! This means that I will have more time to
work on laboratory techniques. It is kind of weird to be finished with something that
seemed so challenging and scary when I first began working in the lab, but I am
excited to move on to the next step in the study. In the meantime, I will likely be
doing more of what I have been doing: slide imaging and cresyl violet staining.
A very exciting opportunity was also presented to me this past week. Dr.
Marla Sunderman, the other postdoc in the lab besides Jenna, has invited me to join
her study for next semester. I have worked with her a couple of times this semester
when she agreed to show me and Jenna some new techniques that could be used in
Jenna’s study. I am very excited about the opportunity, because I had wanted to
participate in the SURF program for the coming summer, but with Jenna’s study
wrapping up in December, I wasn’t sure which study I might want to work with for
the summer. I have a meeting with Marla this next week, so I am hoping to bring up
the possibility of participating in SURF at that time—I hope that her study has SURF
funding! From my experience with Marla, I really like her and think that she would
make a great mentor. I am looking forward to meeting with her and hearing more
about her study, in addition to seeing what my options are for SURF.

11/6/17
For the past couple of weeks, Jenna has not schedule anyone to come into the
lab, since she had a friend’s wedding to go to in Louisiana and then a trip to Europe.
She said that things would be slow anyways since the study is winding down, so she
will not be scheduling anyone until November 17th. Since I have spent significantly
more time in the lab this semester than previous semesters, this is definitely a
welcome break! I do enjoy my time in the lab, but it can definitely be difficult to
juggle with such a challenging class load. So far, the time off is allowing me to sort of
get caught up on studying and life in general.
I had my meeting with Marla, and she went over the premise of her study
with me. She works with cell culture, and she wants to create a new stem cell line
that could be used to study bipolar disorder in rats. I am really excited because her
study sounds so interesting! I also mentioned that I was interested in doing SURF,
and she said that she would see if her lab has funding and let me know, but that she
would love to have me as a student assistant for next semester as well as a SURF
student for the summer. In addition to wanting to continue with research, a big
draw of the SURF program for me is that I want to stay in Cincinnati for the summer
rather than returning home to Akron, as I have done for the past two years. I have
gained so many friends and made so many connections in Cincinnati that it would be
a much more valuable experience for me to stay in Cincinnati for the summer, and
the money provided by SURF would make that all possible. I am really hoping to
hear that SURF funding will be available to me, and in the meantime I will continue
to enjoy a bit of downtime!

11/20/17
I have not been back in the lab in the past two weeks; though Jenna said she
would begin scheduling us on the 17th, she is very sick after returning from her trip
to Europe, so she will not be in the lab to help us with anything we may need until
she is feeling better.
On another note, I have spoken with Marla, and she said that she was told
that SURF funding would be available for me, which is great news! Jenna has also
said that she will write me a letter of recommendation for the program. I am
crossing my fingers that it will all work out for me!

12/4/17
After going home for Thanksgiving, returning to Cincinnati for one week of
class before finals, in addition to being schedule in the lab again, was a big change of
pace. I am happy, however, to be busy again! I am definitely the kind of person who
likes to stay somewhat busy and focused on something, so I am glad to be back in
the lab. I have informed Jenna that although finals are over after this week, I am
willing to stick around for a week if she needs any help with anything, since the
study is wrapping up in a few weeks and she is pretty behind from all of the
setbacks of the last month or so. It looks like I will probably be sticking around until
the 16th, at which point I will head home for the holidays. By the time I return,
Jenna’s study will have fully ended and she will be on her way to her new job in
Memphis, where she will be teaching and doing research at a local college.
I have begun to learn how to do Western blots, and they seem somewhat
complicated, though just like anything else in the lab, they will likely get easier with
practice. It is a two-day process that begins with proteins that have previously been
assayed to ensure that they all have the same weight. They are then inserted into a
gel, which undergoes electrophoresis. The bands on the gel are then transferred to a
membrane, which undergoes lots of washing before and after being tagged with an
antibody. This antibody attaches to certain proteins and causes them to glow when
the membrane is “read” in a special machine, which produces a digital image of the
blot with fluorescence. Though the process does seem a bit complicated, the results
are pretty cool to look at. They can then be analyzed to determine whether or not
certain receptors are present in the samples. I am glad to have learned this
technique, since I have learned in many of my classes that it is a very commonly
used laboratory technique and could translate to other projects that I may work on
in the future.

12/18/17
I am officially finished with the AD-2 study! It is definitely strange to be done
with something that has consumed so much of my time over the past year and a half,
and I am sad to see Jenna go. She has been a mentor and has also become a friend to
me, so it is a bit sad that she will soon be leaving.
 Over the last two weeks I have mainly done proteins assays and western
blots, and they are definitely getting a lot easier! The process seemed sort of lengthy
and frustrating at first, but now that I have nearly gotten the hang of it, I have been
enjoying it and will feel more confident using it in future endeavors.
As for my plans for this next semester and the summer, it seems that there
was a bit of a mistake; Marla’s lab actually does not have funding for any SURF
students this year. It looks like I will have to find a new mentor for the summer;
though I would really like to work with Marla, I do need the money provided by the
SURF program in order to be able to afford to stay in Cincinnati. I have asked around
the lab to see if anyone has any suggestions for who I might try to contact about a
potential SURF position in their lab, but it looks like I will have to do some research
myself and just start looking around at other studies within the hospital and
emailing their PI’s to see if they may be in need of a SURF student for the summer.
Hopefully my previous lab experience will give me a bit of a leg up, in addition to my
recommendation letter from Jenna!
Overall, my experience in the lab has taught me a lot, and I really hope that I
can continue to do research throughout this next semester and summer. I have
definitely gained a lot of valuable experience, and I feel that I have really grown as a
scientist. I am very thankful for the fact that I had such a great mentor who was
always willing to take the time to teach me new things, and I hope that the things
she has taught me will help me as I move forward in the world of scientific research.