Friday, August 18, 2017

Week 11 - 8/14/2017

Week 11

As I wrap up my last week at the lab, it is bittersweet. I'm excited that I've completed my first internship! And I'm excited to go on vacation after this, but I will also miss all of the cool experiments and taking care of the rats.

I am really happy that I decided to intern here because I was able to learn so much and everybody that works there was very helpful to me. Coming into this internship, I knew somewhat what I would be doing but I never thought I would learn as much as I did. I am very thankful to everyone who worked with me this summer because no matter what questions I had they would always be answered. One thing I greatly appreciated about this lab is that everyone works together as a team. Not only does it help everything be more efficient, but it really helps to get everything done correctly and in a timely manner. I am also very happy I got exposure to many different types of experiments and studies. While this internship comes to a close, I am grateful for all the experience I have gained and for all the research work I was able to complete this summer. I can't wait to see where I go next!

Week 10 - 8/7/2017

Week 10

This week’s main focus was working on the attention task. Since all of the Templeton project rats have gone through their two training sessions, we have begun to run them on the A-shape program. They need to complete this program 5 times before they can move onto the 50 signal lever program. We also are still finishing up some of cohort 4 training, and because some of them finished we were able to run a few through the figure 8 locomotive test.

I also spent my time this week weighing and sexing newborn rats for the Diazinon study. This study is a pilot study and is focusing on how the Diazinon pesticide affects development. It is really fun to work with newborns because they are really small and cute. They weigh around eight pounds and by the fourth day weigh around twelve pounds. Since they are little, their eyes are still closed and they don't have any fur yet.

Thursday, August 3, 2017

Week 9 - 7/31/2017

Week 9

This week the main focus was finishing up cohort 4 NICHEs on the attention task and also getting the Templeton project rats acclimated to the attention task program. I detailed in my week 4 blog the different levels of the attention task after acclimation. However, before that the rat has to go through training sessions and the A Shape program.

For the training sessions, the rat is simply put in the box and sugar pellets are dispensed into the food tray. For the first training session, it lasts about 30 minutes. The second training session is a little bit quicker and lasts about 20 minutes. This allows the rats to understand that they are supposed to be trying to get sugar pellets from the food tray. Once the rats have completed those two training sessions, they move on to the A Shape program and have to complete it 5 times.

On Friday, we went to Duke's main campus to see some of the other undergraduate interns at the lab present what they have been working on this summer. We also stopped by the main building for the Duke University Institute for Brain Sciences. It was a fun experience to see other science facilities on Duke's campus.

Weeks 7 & 8 - 7/17/2017 & 7/24/2017

Weeks 7 & 8

For the next two weeks, I ran radial arm maze (RAM) trials. A total of 12 trials were run to see if the animals had learned and had improved their times and memory of going into the correct arm. During the first week, some rats still took the full 10 minutes in the maze, but by the second week all of the rats completed the maze in under 10 minutes. While this was a time consuming experiment because I had to record the data myself and there wasn't a computer doing it like for the attention task, I actually enjoyed it. It was fun to see which rats progressed faster and it was also exciting when a rat would finally understand the maze. There were a few rats that went in the maze in a clockwise or counterclockwise motion, whereas some rats just went to different arms but still finished in under 10 minutes. This was also a cool experiment to see because I have seen the Templeton project rats since they were weaned from their mothers, so it has been interesting to see them "grow up."

For the RAM experiment, the rat is put into the black bucket in the center of the maze for about 10 seconds. The bucket is then removed and the rat explores the arms of the maze and hopefully remembers which ones are baited and which ones are not. Any time the rat enters an arm, it is recorded. At the end of the session, either when the rat has entered all of the baited arms or 10 minutes has passed, the rat is returned to its home cage. Then I calculate the working memory errors, reference memory errors, and latency. Working memory errors are how many times the rat entered an arm the was baited past the initial time it entered, and also if it did not enter a baited arm at all. This is because they are only supposed to enter a baited arm once and remember they entered it. For reference memory errors, that is how many times the rat entered the arms that were not baited. And latency is the time divided by how many times the rat entered each arm.

Radial arm maze

Week 6 - 7/10/2017

Week 6

This week started off slow because we were getting prepared to start the Templetons on another experiment called the Radial Arm Maze. Anytime we start an experiment in a new environment (outside of their home cages) we have to acclimate the rats so that they are used to their surroundings.

The radial arm maze is a 16-arm maze that has 12 out of the 16 arms baited with froot loops. This maze tests two types of memories: working and reference. The purpose is to have the rat remember which arms it is supposed to enter with the froot loops and to not go back into an arm it has already been in. The rats get a reward if they go down the baited arms (a froot loop) and also the quicker they complete the maze the quicker they can go back to their home cage. The time is capped at 10 minutes, so if a rat does not enter all the baited arms they do not get those froot loops. The process for this and working/reference memory will be described in more detail in next week's blog.

For acclimation, the rats needed to get used to the environment and understand that the purpose of being put in this maze was to get froot loops. In order to acclimate them, we put a black bucket in the center of the maze. In the bucket is 12 pieces of froot loops. The rat is then placed in the bucket and taken out once it has eaten all the froot loops or once 10 minutes has passed. This helps the rat understand that when they are put in this environment they are supposed to be looking for the froot loops. 

Radial arm maze

Week 5 - 7/3/2017

Week 5

This week I went back to working with the Templeton study. This week was also a little more complicated than expected because we had July 4th off and then the fire alarm went off on Wednesday during one of our experiments, so it had to be re-done. Although this week was a bit more complicated, it still was a lot of fun to complete some experiments on the Templeton project rats.

The first experiment of the week was novel environment suppressed feeding. In this experiment, the animals were food deprived for 24 hours, and then given food in their cages. When we gave them food, we first weighed the food, set a timer for 10 minutes, and after 10 minutes weighed the food again to see how much they ate. This experiment went smoothly until the fire alarm went off. I was testing two of the last three cages when this happened. Since the fire alarm is really loud and unexpected, it can stress out the animals just like it can stress out humans. So, we were not able to count the testing on those two cages and decided to hold off on testing the last cage since they were most likely stressed too. Since we had to start the experiment over, they needed to be food deprived again for 24 hours. However, since they had just been food deprived we gave them all food to eat for about an hour. Rats normally eat in intervals so this was a sufficient amount of time to make sure they got the nutrition they needed and wanted. They were then food deprived for 24 hours and completed the experiment the next day.

This week was also largely focused on an experiment called "Novel Object Recognition" or as we like to refer to it, NOR. In this experiment, rats are put in a large black box with two of the same objects. An hour later, they are put in the same box with one object from the previous session and a new object. Videos are taken of the rats during both sessions and are scored to see how much time is spent with each object. This experiment tests the memory of the rats. I like this experiment because although it is simple the timing is crucial. Once the rats are done, within the next 10 minutes the videos have to be saved and the boxes have to be cleaned to prepare for the next rats. It is also very important that when the rats are taken out of the boxes the time is recorded because exactly an hour later they have to be put in for their session with the different objects. 

This week was a really fun week because I got to run many experiments on the Templeton project rats. I also enjoyed it because I learned how to deal with the unexpected and adapt to changing schedules.

NOR box
NOR box

Week 4 - 6/26/2017

Week 4

This week I mostly worked on testing the rats in the NICHES study on the attention task. There are four cohorts of rats in total for this study, but the first cohort has finished the attention task. I am currently running cohorts 2, 3, and 4 through this task.

Although it sounds simple, the attention task contains many steps. It is an experiment that tests not only the animal’s ability to pay attention, but also how well it can remember its previous sessions. To start, the rat is trained in the box through a few programs. There are two levers, one left and one right. Depending on which box the rat is in, there is a signal light above the right or left lever. There are multiple stages to this attention task which include: training, A Shape, 50 signal light, 50 blank lever, 50 signal light and 50 blank lever, 100 trials, and 240 trials. I will be taking you through the process of each session below. This is experiment is one that takes the longest because the rat has to pass each program, and if it cannot move on to the next program until it passes. Naturally, this means that some rats are ahead of the others, because just like humans some of them are able to understand the program more easily and quickly.

For the sake of this explanation, I am going to say the signal light is on the left (this also corresponds with the pictures). In the box, the light will go off and if the rat presses the left lever it receives a sugar pellet. Now the rat does not automatically understand this, so the rat goes through training sessions and has to pass them in order to move on. Once it understands that the light means to press the left lever (50 signal light program), it will move on to the program with 50 blank levers, which means that the light will not go off and if it presses the right lever it receives a sugar pellet. Once that program is passed, it goes through the program where it has 50 signals and 50 blanks. Once it completes that program, however, the sessions get more intense.

The next phase that the rat will go through in this attention task is a program we like to refer to as "V." This program is a set of 100 trials with both signals and blanks. Thus, during this if the signal goes off the rat should press the left lever to get a pellet and if it is blank (no light) the rat should press the right lever to get a pellet. However, unlike the previous program this is random so the rat has to pay attention to when the light goes on and when it stays off. This program is slightly more intense because once it passes the regular "V" session we move the signal light farther up to the "V+1" position. The rat then goes through 100 trials, and has to pass. A passing score is a score greater than or equal to 70%. The signal light then gets moved up one more spot, and the rat again has to pass 100 trials in the "V+2" session. The third and final phase of this "V" program is moving the light into the center of the box. Again, the rat has to pass 100 trials in this program we refer to as "V+C." Once that is passed, the rat is almost done with the attention task as it has one more phase to go through. 

The last and final phase of the program is what we refer to as "V7." The signal light is in the center of the box, like the "V+C" position, but the rat has to pass 240 trials this time. Now, unlike the other programs, the rat has to pass the “V7” program six times. Once it has passed six times, the rat has successfully completed the attention task.

While the program itself can take time, we also have to always make sure the boxes are working correctly. Sometimes the sugar pellets will get stuck to the metal plates they come down from, so we have to make sure those are clean. We also regularly clean out the bottom of the cages. Although the attention task was a little daunting at first, I have come to really enjoy it. I started working on this in the middle of cohort 2 and 3 completing the task, and when cohort 4 was just going through the training sessions. It is really enjoyable and fun to see which rats move through the programs quickly and it is fun to see which rats pass! Overall, this week was pretty enjoyable because I was also able to run some of the NICHES rats through figure 8 training. I am really starting to have fun with the work I am doing as I am getting more involved with the experiments. 

Attention task box
Attention task box with signal light at V+C and V7 position

Week 3 - 6/19/2017

Week 3

During my third week I was a lot more involved with the testing of the rats. I worked with the Templeton project rats for the figure 8 maze and the NICHES on the attention task, which will be detailed in next week’s blog. My main focus this week will be to explain the figure 8 locomotive test I completed on the Templeton project rats.

I have used programs before to analyze statistics, but I have never used programs to conduct experiments so I thought the whole process was pretty neat. In this program, a case file has to be created and the rat ID's have to be matched to the box I would load them into. This is the most intensive part of this program because I have to make sure the ID’s match the correct box as this is how we are able to track the data of the rat.

While the program is pretty old, the boxes are in good condition. Like the name of the test entails, it is a figure 8 maze that has sensors throughout that track the rat’s movement. Once I setup the program, I place the rats in the figure 8 maze for an hour and then take them out. Recording the data is a more precarious task because there are quite a few steps that need to be taken in order to get the data, and then the data has to be recorded on a sheet and then put in a folder on the computer. Once that is done, I transfer the data from my sheet onto an excel sheet with the rest of the experimental data on an up-to-date computer. This task is one of my favorites to complete because I enjoy being able to work with the computer and program, even if it can be difficult at times and is older.

I also think this test is really fascinating because it is able to assess whether or not the rats are hyperactive and have ADHD or ADD. This week definitely felt amazing as I really felt I had made my first major contribution to helping out with research! I am really looking forward to what the next weeks have in store for me.

Figure 8 maze boxes
Figure 8 maze box
Figure 8 maze box open

Week 2 - 6/12/2017

Week 2

As my second week began, I knew it was about to get a lot more interesting because I was added on protocol. This meant that I was now able to handle the animals. At the Behavioral Research Lab they conduct multiple studies on the rats and fish they have, but my time here will be focused on two main projects.

The first project is the NICHES study, and details about the study can be found here. NICHES stands for the study of "neurodevelopment and improving children's health following environmental tobacco smoke exposure." Currently with this project we are running the rats through a program called the attention task (more details to follow in next week’s blog) and then studying their sexual behavior and neurotoxicology.

The second project is the Templeton study, which is a study of how THC affects epigenetics. We are currently studying offspring of males who were exposed to THC orally. We are running them through multiple behavioral tests. Again, more details on the tests and progress of the study will be detailed in upcoming weekly blogs.

Although I was added to protocol this week, a lot of what I did was basic and simple things to make sure the rats were being taken care of. I spent a few days making cages, changing cages, marking the rat's tales so we can tell which rat is which, weighing them, feeding them, and checking on them. I realized that no matter where you go in life you are going to need to complete tasks that you do not always want to because they seem mundane, boring, or not important enough. But without all of these simple tasks completed, not only do the rats not live good lives, but no testing would be able to be completed. Millions of lives would not be able to be helped with research just because I didn't want to do something simple and easy. So even though it was boring at points to just be changing cages, I knew it was just as important as running tests. I did get to run one test this week--it is called an elevated plus-maze. I completed this task with one fellow assistant. During the task we timed when the rats would stay in the open ends, when they stay in the closed ends, and when they cross from open arm to open arm or closed arm to closed arm. All in all, even though this week wasn't what I expected, I still learned a lot and obtained a greater appreciation for all of the research that is completed throughout the world. 

The elevated plus maze
The elevated plus maze

Friday, June 30, 2017

Week 1 - 6/5/2017

Week 1

Me standing in front of the lab building and sign
The door to the lab
The wet lab
The pH scale machine
The view of the lab from across the street

It was the first week of my internship and I was not sure what to expect. I had just finished my first year of college, and had taken about a two week break to get settled at home. I have never had a 9-5, Monday-Friday job before, so this was new territory for me. I knew that my Magellan would be a lot different from many others--after all I am staying in the United States. But I also knew that it would be a worthwhile experience. Already in my first week I was exposed to many lab practices, protocols, rules, and regulations that I had never thought of before.

During the first week, I went through training about animal care, how to handle animals, laboratory safety, and the ethics of animal testing. The lab that I am working with completes studies on rats and fish. I had not been added to protocol yet, so a lot of my first week was watching what the lab technicians do and helping out with small tasks like making cages. To me, this was the perfect way of settling into my job as a lab assistant because I really got to observe the method and type of work the technicians did.

The two most fascinating parts of my week was making nicotine for the self-administration rats and being able to dissect rats for toxicology studies. I made the nicotine in the lab alongside a fellow assistant after being taught once and walked through it by one of the lab technicians. I really enjoyed this because I felt like I was finally being able to apply what I had learned in my chemistry labs to something useful in the work world. We had to test the pH of the solution and adjust it to be just right by using NaOH and HCl solutions. It was really fun because I was able to use a digital pH detector instead of paper!

It also enjoyed learning how to dissect the spleen and brain out of rats, because I took an anatomy and physiology class in high school and dissected cats. At first it was a little bit tricky, but after observing some dissections and being walked through a few, I got the hang of it.

My first week was definitely very rewarding, because I learned a lot about how labs operate and all of the protocols and guidelines that go into research. I was also happy the I was able to survive my first week of work in the adult world! I know that I will learn a lot more throughout this internship and I am very grateful for the opportunity to complete it.