So I have officially left the lab =(
The last couple of days Dr. Cotten was back from the OR. My ligation finally did work at the end of last week, so we grew up some of the colonies. Out of the 18 colonies I picked, two of them looked promising, so we sent them off for sequencing.
Since Dr. Cotten was around and such, in the time between steps in the experiments, he taught me more things about the lab and what it was doing, as well as showing me how they test to see if the RNA we make from the DNA template of our gene is actually expressing.
See the gene we're working on acts on IN channels. These are transmembrane proteins that are specific to Potasium, meaning that the only thing that can get through the membrane via this protein is Potasium. We inject oocytes (frog eggs!) with the RNA, and hook it up to a machine to see if it is expressed. We can tell if it is expressing by running a current through the oocyte and seeing if the potasium gradient changes when we clamp the voltage at a specific spot.
So, in summary, I have really enjoyed the time I have spent in this second lab. I would have prefered to have had more time in it so I could really have gotten more in-depth with everything, but you make do with what life gives you! As it turns out I will not be going back to the lab later (I need time to go visit colleges! And maybe just a couple days I can kick back and relax...) but Dr. Cotten said he would keep me updated with some of the continuing research he's doing, continuing on from what I had been doing with him.
~Charlotte <3
Sunday, August 9, 2009
Wednesday, July 29, 2009
Weeks 5 and 6
So I haven't posted in a while!
For the past two weeks I have been in the Edwards Research Lab on the main campus of MGH, and I am enjoying it a lot more than when I was in the Navy Yard (although I do miss the building).
For the first week, Dr. Cotten showed me the ropes and tried to teach me to do the basics on my own- DNA purification, running gells... things like that. I tried to absorb as much as possible, and - considering the short amount of time I had - I think I did pretty well doing things on my own.
When Monday rolled around and Dr. Cotten was in the OR, however, it was a different story. I was completely on my own, and although I could call Dr. Cotten or page him if I needed help, there were tons of little questions I had that weren't worth bothering him with that I had to figure out on my own. I think I managed to do everything alright, though! Not that I always got the right results (or any results at all...).
So here is my daily routine (although it changes from day to day depending on what needs to be done). I wake up at 8, am at breakfast in Annenburg dinning hall at Harvard by 8.30, pack my lunch so I don't need to buy it, and go to the T. I am usually at work by 9.15-9.30. Then I usually have an email from Dr. Cotten with instructions as to what to do for the day while he's in the OR. I get to work, and while things are incubating or mixing (usually in one-hour increments) I either read for psychology, or busy myself with whatever might be in my backpack. I have a couple papers from Dr. Cotten that he gave me the first week that I've been trying to work through, as well as my summer spanish reading for class this fall. Dr. Cotten usually tries to swing by during his lunch break (between 12 and 1) to check up on me. Then I go eat lunch outside on the green right outside the lab's window (well, 5 stories below), and come back and do more experiments in the afternoon. Usually I am out of here around 4, although yesterday I didn't leave until 5.30, because Dr. Cotten got out early from the OR and did a digest to check we had the right DNA.
The past two days I did exactly the same thing, because I was supposed to be ligating digested DNA from last friday and inserting it into ecoli to grow up. Both yesteday morning and this morning I came in to find no colonies on my plates, which means I've had to do some troubleshooting. Since we ran out the DNA to be sure we had all the parts to it yesterday just to check, I know there's nothing wrong with the DNA I started off with myself, so the problem is probably that somehow the DNA is not getting into the ecoli. As it has failed twice, Dr. Cotten has ordered some different ecoli to see if that will work. They may be too old or something.
So this morning I don't think I have much to do, as I can't do the next step until the new ecoli shows up, and Dr. Cotten didn't give me any further instructions. The ncie thing is that I have this lab at my disposal, so I will probably putter around and see if I can't just make sure there isn't something wrong with a different stage of the process and somehow I am messing up the DNA.
Dr. Cotten also had another idea for a project, involving trying to attach another gene to his dimer, which codes for a protein that will apmlify the effects of his first protein by 5-fold or so. I think he was planning on doing it with me next week when he's out of the OR, but I have to leave on Tuesday evening so I can get back home for my music camp on Wednesday. So I may end up coming back to this lab after my music camp is over (If I'm not completely and utterly exhausted) and doing this project with him then. I would be really excited! The only thing is that it will deprive me of a little more of my summer, which has already not been a vacation and since I will be spending the majority of my free time doing college app prep, it doesn't look like I am going to get much of one!
For the past two weeks I have been in the Edwards Research Lab on the main campus of MGH, and I am enjoying it a lot more than when I was in the Navy Yard (although I do miss the building).
For the first week, Dr. Cotten showed me the ropes and tried to teach me to do the basics on my own- DNA purification, running gells... things like that. I tried to absorb as much as possible, and - considering the short amount of time I had - I think I did pretty well doing things on my own.
When Monday rolled around and Dr. Cotten was in the OR, however, it was a different story. I was completely on my own, and although I could call Dr. Cotten or page him if I needed help, there were tons of little questions I had that weren't worth bothering him with that I had to figure out on my own. I think I managed to do everything alright, though! Not that I always got the right results (or any results at all...).
So here is my daily routine (although it changes from day to day depending on what needs to be done). I wake up at 8, am at breakfast in Annenburg dinning hall at Harvard by 8.30, pack my lunch so I don't need to buy it, and go to the T. I am usually at work by 9.15-9.30. Then I usually have an email from Dr. Cotten with instructions as to what to do for the day while he's in the OR. I get to work, and while things are incubating or mixing (usually in one-hour increments) I either read for psychology, or busy myself with whatever might be in my backpack. I have a couple papers from Dr. Cotten that he gave me the first week that I've been trying to work through, as well as my summer spanish reading for class this fall. Dr. Cotten usually tries to swing by during his lunch break (between 12 and 1) to check up on me. Then I go eat lunch outside on the green right outside the lab's window (well, 5 stories below), and come back and do more experiments in the afternoon. Usually I am out of here around 4, although yesterday I didn't leave until 5.30, because Dr. Cotten got out early from the OR and did a digest to check we had the right DNA.
The past two days I did exactly the same thing, because I was supposed to be ligating digested DNA from last friday and inserting it into ecoli to grow up. Both yesteday morning and this morning I came in to find no colonies on my plates, which means I've had to do some troubleshooting. Since we ran out the DNA to be sure we had all the parts to it yesterday just to check, I know there's nothing wrong with the DNA I started off with myself, so the problem is probably that somehow the DNA is not getting into the ecoli. As it has failed twice, Dr. Cotten has ordered some different ecoli to see if that will work. They may be too old or something.
So this morning I don't think I have much to do, as I can't do the next step until the new ecoli shows up, and Dr. Cotten didn't give me any further instructions. The ncie thing is that I have this lab at my disposal, so I will probably putter around and see if I can't just make sure there isn't something wrong with a different stage of the process and somehow I am messing up the DNA.
Dr. Cotten also had another idea for a project, involving trying to attach another gene to his dimer, which codes for a protein that will apmlify the effects of his first protein by 5-fold or so. I think he was planning on doing it with me next week when he's out of the OR, but I have to leave on Tuesday evening so I can get back home for my music camp on Wednesday. So I may end up coming back to this lab after my music camp is over (If I'm not completely and utterly exhausted) and doing this project with him then. I would be really excited! The only thing is that it will deprive me of a little more of my summer, which has already not been a vacation and since I will be spending the majority of my free time doing college app prep, it doesn't look like I am going to get much of one!
Tuesday, July 14, 2009
New Lab!
Wow. What a roller coaster.
So I was a little wound up because I didn't have things to occupy my time and I was worrying about what to to from day to day, and I was feeling pretty lonely at Harvard some of the time. On top of that, I'm sick, and I haven't been able to get a good nights sleep because all my roommates are out until who knows what hour and are going in and out of the room a lot starting around 9pm when I have just decided it's not too early to hop into bed.
So yesterday, I ended up leaving the lab around 2.30. Dr. Mao hadn't called yet, but I asked Calvin if he would talk to him for the both of us. He knew I was sick, and I think I must have looked like a real wreck because he said not to worry about it and I should really go home and get some sleep so I would feel better.
I went over to Dr. Dunn's office and explained about everything that had gone on at the lab and what a hard time I was having there coupled with being away from home. I felt horrible because I felt as though I was complaining; I had been given this incredible opportunity to be in a research lab with really cutting edge research for the summer, and here I was going on to the guy had organized it about what a hard time I'm having with his program. Dr. Dunn was so nice about everything though, and made me feel so much better about it. He said he had had an alternative worked out at the beginning of the summer in case this lab didn't work out, because it was the first time SFS had sent a student over there. So he called up the guy in the alternative lab, and even before he had sent the email to both of us as the introduction, the other doctor had emailed me and given me his cell phone number and suggested we meet sometime the next day. And even then, if this new lab didn't look like it was going to work out, and there wasn't anything for me to do there, then I could come right back to him and he would do his best to find another lab for me.
Today I met with Dr. Cotten. He's the person from the new lab that Dr. Dunn had been in touch with. I thought I was just going to be given an overview of what I would be doing and the people in the lab and such, but I ended up being there until around 4pm! Dr. Cotten is extremely nice, and he explained all about what the purpose of the lab is and his focus of research, and then went into details about what I specifically would be doing. It was almost like I was back in AP bio, except that the class only included me (although there was a white board and he was using it!) and the teacher looked remarkably more like Mr. Palm than Dr. K (seriously, if you look at him from the side he looks just like him!). I understood almost all of what he was explaining; it was SO much more relevant to the material I learned in AP bio (much more molecular-based) than things in the other lab had been, as they had mostly been about anatomy. I'll elaborate more when I have a more solid understanding of exactly what it is I'm doing, but long story short, most of what I am going to be doing is working with e-coli! Except these colonies don't glow in the dark like the ones I did in AP bio class, so it's more difficult to identify which ones are the colonies with our gene transformed into plasmids in the bacteria. So the project is mostly cell culturing as opposed to slicing and dicing rats that have been forced to swim for long periods of time.
We made the LB Augur, which goes on the bottom of the plates so the bacteria can grow and replicate, as well as pick the lone four colonies that has grown from Dr. Cotten's previous sample. We're not sure it's going to work out because this sample is 5 years old, but we'll see.
So the lab is on the main campus of MGH (which is a shame, really, because although my commute will be shorter, I really liked being in the Navy Yard building; it is such a wonderful place...) and I'm pretty sure I know how to get to it form the main entrance now! I've been having difficulty finding my way around MGH... it's just so BIG! This lab is smaller and more cluttered than the lab at the Navy Yard, but it seems a bit more functional that way- like there's more going on, or that there's more that COULD be going on.
And I will have my own things to be doing! When I was at the Navy Yard there were a couple of interns (who looked like they were in college) who were working in the lab next to mine, and they were constantly occupied, and half the time I'd walk by them and different people in their lab would be explaining to them what to do. I feel like I will be functioning much more like them now than I had been. A lot of my time at the Navy Yard was just spent holed up in our little "office" sending emails or reading to pass the time.
The only problem is that Dr. Cotten is going to be in the OR for the next two weeks, because he's tutoring a medical intern. He said he was going to try to pick cases that were earlier in the morning so he may be able to get out before I would leave and help me along, but I am really going to be doing a lot of this by myself. I was really nervous about this at the beginning of the day, but I've got tomorrow and Friday to get the hang of things, and I'm going to try to get an outline written down just so I have it to refer to so I know what the next step in my process will be. And he introduced me to another couple of people in the lab who I can go to for help. In particular, there is one girl who is at the lab bench next to his who's a graduate student. She has an intern for the summer too, who she'll be showing the ropes to, so I can watch her do things and ask for help from her if I need it. And Dr. Cotten said he will try to be in touch as much as possible to help me along. I've got his cell number, and I can always page him (though I will have to remember to get his page number tomorrow).
So I think I will definitely be switching to this new lab. I was worried about not having an anchor, and it's true that I don't have my own sort of "space" in this lab other than Dr. Cotten's workbench, but as he isn't going to be there most of the time anyway, I guess I kind of do. I think I will also sack the whole commuting idea (I was toying with it earlier when people started saying they were going to switch me around places, so that I could handle the transition better).
So I was a little wound up because I didn't have things to occupy my time and I was worrying about what to to from day to day, and I was feeling pretty lonely at Harvard some of the time. On top of that, I'm sick, and I haven't been able to get a good nights sleep because all my roommates are out until who knows what hour and are going in and out of the room a lot starting around 9pm when I have just decided it's not too early to hop into bed.
So yesterday, I ended up leaving the lab around 2.30. Dr. Mao hadn't called yet, but I asked Calvin if he would talk to him for the both of us. He knew I was sick, and I think I must have looked like a real wreck because he said not to worry about it and I should really go home and get some sleep so I would feel better.
I went over to Dr. Dunn's office and explained about everything that had gone on at the lab and what a hard time I was having there coupled with being away from home. I felt horrible because I felt as though I was complaining; I had been given this incredible opportunity to be in a research lab with really cutting edge research for the summer, and here I was going on to the guy had organized it about what a hard time I'm having with his program. Dr. Dunn was so nice about everything though, and made me feel so much better about it. He said he had had an alternative worked out at the beginning of the summer in case this lab didn't work out, because it was the first time SFS had sent a student over there. So he called up the guy in the alternative lab, and even before he had sent the email to both of us as the introduction, the other doctor had emailed me and given me his cell phone number and suggested we meet sometime the next day. And even then, if this new lab didn't look like it was going to work out, and there wasn't anything for me to do there, then I could come right back to him and he would do his best to find another lab for me.
Today I met with Dr. Cotten. He's the person from the new lab that Dr. Dunn had been in touch with. I thought I was just going to be given an overview of what I would be doing and the people in the lab and such, but I ended up being there until around 4pm! Dr. Cotten is extremely nice, and he explained all about what the purpose of the lab is and his focus of research, and then went into details about what I specifically would be doing. It was almost like I was back in AP bio, except that the class only included me (although there was a white board and he was using it!) and the teacher looked remarkably more like Mr. Palm than Dr. K (seriously, if you look at him from the side he looks just like him!). I understood almost all of what he was explaining; it was SO much more relevant to the material I learned in AP bio (much more molecular-based) than things in the other lab had been, as they had mostly been about anatomy. I'll elaborate more when I have a more solid understanding of exactly what it is I'm doing, but long story short, most of what I am going to be doing is working with e-coli! Except these colonies don't glow in the dark like the ones I did in AP bio class, so it's more difficult to identify which ones are the colonies with our gene transformed into plasmids in the bacteria. So the project is mostly cell culturing as opposed to slicing and dicing rats that have been forced to swim for long periods of time.
We made the LB Augur, which goes on the bottom of the plates so the bacteria can grow and replicate, as well as pick the lone four colonies that has grown from Dr. Cotten's previous sample. We're not sure it's going to work out because this sample is 5 years old, but we'll see.
So the lab is on the main campus of MGH (which is a shame, really, because although my commute will be shorter, I really liked being in the Navy Yard building; it is such a wonderful place...) and I'm pretty sure I know how to get to it form the main entrance now! I've been having difficulty finding my way around MGH... it's just so BIG! This lab is smaller and more cluttered than the lab at the Navy Yard, but it seems a bit more functional that way- like there's more going on, or that there's more that COULD be going on.
And I will have my own things to be doing! When I was at the Navy Yard there were a couple of interns (who looked like they were in college) who were working in the lab next to mine, and they were constantly occupied, and half the time I'd walk by them and different people in their lab would be explaining to them what to do. I feel like I will be functioning much more like them now than I had been. A lot of my time at the Navy Yard was just spent holed up in our little "office" sending emails or reading to pass the time.
The only problem is that Dr. Cotten is going to be in the OR for the next two weeks, because he's tutoring a medical intern. He said he was going to try to pick cases that were earlier in the morning so he may be able to get out before I would leave and help me along, but I am really going to be doing a lot of this by myself. I was really nervous about this at the beginning of the day, but I've got tomorrow and Friday to get the hang of things, and I'm going to try to get an outline written down just so I have it to refer to so I know what the next step in my process will be. And he introduced me to another couple of people in the lab who I can go to for help. In particular, there is one girl who is at the lab bench next to his who's a graduate student. She has an intern for the summer too, who she'll be showing the ropes to, so I can watch her do things and ask for help from her if I need it. And Dr. Cotten said he will try to be in touch as much as possible to help me along. I've got his cell number, and I can always page him (though I will have to remember to get his page number tomorrow).
So I think I will definitely be switching to this new lab. I was worried about not having an anchor, and it's true that I don't have my own sort of "space" in this lab other than Dr. Cotten's workbench, but as he isn't going to be there most of the time anyway, I guess I kind of do. I think I will also sack the whole commuting idea (I was toying with it earlier when people started saying they were going to switch me around places, so that I could handle the transition better).
Monday, July 13, 2009
Hello...? Anyone here...?
When I left the lab on Wednesday, I told the people in my lab that I was going, and that I wouldn't be there the next day because I was shadowing a doctor at MGH, so I'd be back and see them on Friday.
I had an awesome time on Thursday. I followed an anesthesiologist who has been working there for 6 years. She showed me around the OR and gave me an overview of MGH as a whole so I wouldn't get lost, and was explaining things all the time. I saw one woman go under before her surgery, and then the doctor left me with another anesthesiologist. He was doing the anesthesia for a patient who had a spinal tumor. Apparently, this is usually a fairly routine procedure. The only difference with this one was, that the patient was rather overweight, so they had much more difficulty doing the simple things that prepared for the operation. It was really interesting. I left around noontime, though, because the doctor I had been following originally had been going on about how it was the first nice day for such a long time and I should take the afternoon off and go "enjoy youth." I also can't get back into the OR without another doctor, so I actually would have had difficulty trying to get back into the OR after having had lunch. She is going on vacation for a week so I can't follow her next week, but she should be back the week after. We'll see!
I don't mind when people are lazy, but it would be nice if they would just let me know when they are going to be lazy so I can avoid wasting my time. When I came in on Friday, after not having been there for a day, Neither Shuxiang nor Connie were there. I figured they would probably just be coming in late, or either they were in a different room somewhere, so I went back to my "office" and occupied myself, going back to check if they were there in 15 minute intervals. Calvin was there too, but he had some testing he had to do in the afternoon for Pecure while he was in Africa (Calvin is following Pecure, and Pecure went to Africa for a conference for two weeks, so Calvin in doing some of his research for him while he's away). Anyway, after about an hour and a half of this, I remembered I had Connie's cell phone number, so I texted her and asked her where she was. She said she was feeling lazy and might come into the lab in the afternoon. I asked her what I should do because Shuxiang wasn't there, and she said I could probably just go home. So I wasted my whole morning and woke up early for nothing, pretty much. It just would have been nice if they had let me know they weren't coming in the next day, because they haven't given me anything to do that I can do without them.
Then I came in this morning (the next Monday), expecting to start my project (because I thought the rats were coming in today), to find that they're actually coming tomorrow. Calvin isn't going to be here tomorrow, and since we're doing the project together, that means we can't start until Wednesday. So then we decided we'd go talk to Dr. Mao and straighten some details out, but he wasn't there, so we asked Shuxiang to page him. He was busy, so he is apparently going to call back this afternoon. I asked Shuxiang if there was anything I could do for him today (like clean his samples if he didn't feel like doing it or something), and he said he was going to clean his samples in 45 minutes and then incubate them for the night, and that was it. So there wasn't that much going on today.
So, I am stuck occupying myself all day until Dr. Mao calls back (Calvin too, actually), and then I will ask Shuxiang if there is something I can actually do tomorrow, and if there isn't, then I think I won't bother coming in, because I'll just be sitting in my little side room all day. doing stuff that I could be doing back in my dorm room at Harvard, or at home for that matter.
I will probably be able to start my own project on Wednesday, then I will be in the hospital on Thursday, and back in the lab doing my project again on Friday. I just have to figure out what to do with the rest of today, and Tuesday. The only thing is... This is exactly what I was doing last week... thinking that on Monday I wouldn't have to worry about what I would fill my time with anymore...
I had an awesome time on Thursday. I followed an anesthesiologist who has been working there for 6 years. She showed me around the OR and gave me an overview of MGH as a whole so I wouldn't get lost, and was explaining things all the time. I saw one woman go under before her surgery, and then the doctor left me with another anesthesiologist. He was doing the anesthesia for a patient who had a spinal tumor. Apparently, this is usually a fairly routine procedure. The only difference with this one was, that the patient was rather overweight, so they had much more difficulty doing the simple things that prepared for the operation. It was really interesting. I left around noontime, though, because the doctor I had been following originally had been going on about how it was the first nice day for such a long time and I should take the afternoon off and go "enjoy youth." I also can't get back into the OR without another doctor, so I actually would have had difficulty trying to get back into the OR after having had lunch. She is going on vacation for a week so I can't follow her next week, but she should be back the week after. We'll see!
I don't mind when people are lazy, but it would be nice if they would just let me know when they are going to be lazy so I can avoid wasting my time. When I came in on Friday, after not having been there for a day, Neither Shuxiang nor Connie were there. I figured they would probably just be coming in late, or either they were in a different room somewhere, so I went back to my "office" and occupied myself, going back to check if they were there in 15 minute intervals. Calvin was there too, but he had some testing he had to do in the afternoon for Pecure while he was in Africa (Calvin is following Pecure, and Pecure went to Africa for a conference for two weeks, so Calvin in doing some of his research for him while he's away). Anyway, after about an hour and a half of this, I remembered I had Connie's cell phone number, so I texted her and asked her where she was. She said she was feeling lazy and might come into the lab in the afternoon. I asked her what I should do because Shuxiang wasn't there, and she said I could probably just go home. So I wasted my whole morning and woke up early for nothing, pretty much. It just would have been nice if they had let me know they weren't coming in the next day, because they haven't given me anything to do that I can do without them.
Then I came in this morning (the next Monday), expecting to start my project (because I thought the rats were coming in today), to find that they're actually coming tomorrow. Calvin isn't going to be here tomorrow, and since we're doing the project together, that means we can't start until Wednesday. So then we decided we'd go talk to Dr. Mao and straighten some details out, but he wasn't there, so we asked Shuxiang to page him. He was busy, so he is apparently going to call back this afternoon. I asked Shuxiang if there was anything I could do for him today (like clean his samples if he didn't feel like doing it or something), and he said he was going to clean his samples in 45 minutes and then incubate them for the night, and that was it. So there wasn't that much going on today.
So, I am stuck occupying myself all day until Dr. Mao calls back (Calvin too, actually), and then I will ask Shuxiang if there is something I can actually do tomorrow, and if there isn't, then I think I won't bother coming in, because I'll just be sitting in my little side room all day. doing stuff that I could be doing back in my dorm room at Harvard, or at home for that matter.
I will probably be able to start my own project on Wednesday, then I will be in the hospital on Thursday, and back in the lab doing my project again on Friday. I just have to figure out what to do with the rest of today, and Tuesday. The only thing is... This is exactly what I was doing last week... thinking that on Monday I wouldn't have to worry about what I would fill my time with anymore...
Tuesday, July 7, 2009
Week Three...
I shall summarize what I have been doing so far.
The first two days I did very little. I watched people do a behavior test and then cut spinal chord samples the next day. I think the main problem was that I didn't know quite what I was here for in the first place, and was expecting them to give me something to do and keep doing for them, but instead of that, the people in my lab didn't seem to know quite what to do with me either, so I ended up kind of just staying in the little room they provided for me to put my stuff, waiting for them to come knock on the door and tell me to do something.
Then the next day Connie took me under her wing and showed me how to do immunohistochemistry for the remainder of the first week. This is basically "tagging" of proteins of interest. We used immunofluorescence, so the antibodies we used to tag the antigens we were interested in were fluorescent. Once we had finished tagging them on Friday, we took a bunch of pictures of them with this microscope that is connected to the computer so we can capture images easily. I going to try to attach a couple of them... If i can figure out how to do it (blogs are new to me)
The next week Connie taught me how to do Western Blotting. Wow. Who'd have thought research could be so tiring!? There were a million steps and it really did take all week.
The first step was to make the gel. I did a decent job, I think. It was a lot thinner than the gels I remember running in AP Bio, and we had to make a separating gel first, and then a stacking gel to put on top of it, which forms the wells where the samples go in.
The second step was to do a protein assay. I honestly don't remember all the specifics, although I have written them all down in my lab notebook, but we had to make a standard curve first and read the protein concentrations of those samples, which functioned as controls, and then read the concentrations of our actual samples.
The protein we were interested in for this one was BDNF (brain-derived neurotrophic factor), and we had to prepare the samples and then run the gels.
Once we had done that, we went through the process of blocking (I think), and moved the proteins from the gels onto a transferable membrane, which (to my eyes) was just a really thin piece of special paper. Then we did a similar kind of thing as we did in immunofluorescence, washing the filter papers in PBS (Phosphate Saline Buffer, which is supposed to keep the pH of the sample stable) and then adding the antibodies to tag the protein we were interested in. This time, however, the antibodies were radioactive, instead of fluorescent. Thus, when we had finished tagging them, we took the samples to the dark room and copied them onto film paper. I got to keep the results as a "souvenir" or so Connie called it.
The problem with all this, is that Connie took time out of her day and her research to teach me how to do these tasks, when, technically, I am supposed to be following Shuxiang. He is more difficult to understand than Connie, though. I didn't really know what to expect coming into this, but I did think I was supposed to be being useful in some way, instead of taking up someone's time. I thought Connie was pretty much done with her research, because she is only here for a year doing research for her PhD, and she gave a summary presentation last Monday on her findings, but yesterday she was back to doing her own research, as she was cutting up brain samples and tagging her own protein: NF-kappa-B.
Yesterday, I followed Calvin as he was watching Pecure (I think that's how you spell his name), the researcher he's following, taking samples from his rats. I don't know if I'm technically supposed to disclose information about exactly what he was doing, but he also let Calvin and me try, and I have to say, it was pretty disgusting. I really don't like doing it and I am going to try to avoid ever doing it again.
Yesterday morning we were supposed to have a lab meeting, but the person who was going to be presenting hadn't finished her slides, so Calvin and I went to meet with Dr. Mao instead. He started by doing a basic overview of the purpose of his lab, and why he does both research and Clinical work for his job. Then he basically laid the table open and said that Calvin and I could either write a review, or do a mini-project of our own. If we wrote a review, basically what we would do is do a lot of reading and summarize something about the biology relating to what is being done in the lab. For example, we would do a bunch of research on the biology of a spinal chord, for example, because that would come in handy when trying to understand what all the images people were looking at of spinal chords meant with the differing expressions in proteins, etc, etc. But I'd really rather not just do a bunch of reading and then write a paper, because, firstly, I could do that anywhere, I don't need to be in a research lab, and secondly, it's the middle of the summer and I have written plenty of paper's this past year. Calvin was on the same page as me, so we decided we would figure out something we could do for a project. We went back to Dr. Mao a bunch of times with different ideas, and eventually we came up with a plan.
Our mini-project is going to be this: We want to know the effect on pain thresholds of rats when induced with depression by forced swim tests. This should take me to the end of July.
The problem left to me now is what to do with this week. I have 3 days left of work this week and nothing planned for me, that I know of. If they don't have anything for me to do I suppose I will research more into what the results of our experiment could mean, or I could always occupy myself by reading for my psych class.
Today, for example, I got here a bit late because I had to get a weekend slip from Harvard and the office doesn't open until 9am. Then I did a bit of research and wrote this post, and when I went to find Shuxiang, he said that he wasn't doing anything of interest today, just writing his paper, I think (again, it was hard to understand him). He can't continue with his research until a specific antibody comes in, which he has been waiting for for two weeks because it's on back-order. And they were done with the behavioral testing for the day, and won't do it again until Thursday, so I can't go watch it. So basically I am going to go eat lunch now, and then go back to Harvard, at 1pm. If I was a slacker and liked not having to do anything, this would be fabulous. But as it is, I'm just going to end up reading Harry Potter all afternoon, which I guess is fine, but I'm supposed to be at work... So I can't say I'm happy about it, really.
I get to shadow a doctor through the OR in MGH on Thursday, though! I'm really excited about that!!
The first two days I did very little. I watched people do a behavior test and then cut spinal chord samples the next day. I think the main problem was that I didn't know quite what I was here for in the first place, and was expecting them to give me something to do and keep doing for them, but instead of that, the people in my lab didn't seem to know quite what to do with me either, so I ended up kind of just staying in the little room they provided for me to put my stuff, waiting for them to come knock on the door and tell me to do something.
Then the next day Connie took me under her wing and showed me how to do immunohistochemistry for the remainder of the first week. This is basically "tagging" of proteins of interest. We used immunofluorescence, so the antibodies we used to tag the antigens we were interested in were fluorescent. Once we had finished tagging them on Friday, we took a bunch of pictures of them with this microscope that is connected to the computer so we can capture images easily. I going to try to attach a couple of them... If i can figure out how to do it (blogs are new to me)
The next week Connie taught me how to do Western Blotting. Wow. Who'd have thought research could be so tiring!? There were a million steps and it really did take all week.
The first step was to make the gel. I did a decent job, I think. It was a lot thinner than the gels I remember running in AP Bio, and we had to make a separating gel first, and then a stacking gel to put on top of it, which forms the wells where the samples go in.
The second step was to do a protein assay. I honestly don't remember all the specifics, although I have written them all down in my lab notebook, but we had to make a standard curve first and read the protein concentrations of those samples, which functioned as controls, and then read the concentrations of our actual samples.
The protein we were interested in for this one was BDNF (brain-derived neurotrophic factor), and we had to prepare the samples and then run the gels.
Once we had done that, we went through the process of blocking (I think), and moved the proteins from the gels onto a transferable membrane, which (to my eyes) was just a really thin piece of special paper. Then we did a similar kind of thing as we did in immunofluorescence, washing the filter papers in PBS (Phosphate Saline Buffer, which is supposed to keep the pH of the sample stable) and then adding the antibodies to tag the protein we were interested in. This time, however, the antibodies were radioactive, instead of fluorescent. Thus, when we had finished tagging them, we took the samples to the dark room and copied them onto film paper. I got to keep the results as a "souvenir" or so Connie called it.
The problem with all this, is that Connie took time out of her day and her research to teach me how to do these tasks, when, technically, I am supposed to be following Shuxiang. He is more difficult to understand than Connie, though. I didn't really know what to expect coming into this, but I did think I was supposed to be being useful in some way, instead of taking up someone's time. I thought Connie was pretty much done with her research, because she is only here for a year doing research for her PhD, and she gave a summary presentation last Monday on her findings, but yesterday she was back to doing her own research, as she was cutting up brain samples and tagging her own protein: NF-kappa-B.
Yesterday, I followed Calvin as he was watching Pecure (I think that's how you spell his name), the researcher he's following, taking samples from his rats. I don't know if I'm technically supposed to disclose information about exactly what he was doing, but he also let Calvin and me try, and I have to say, it was pretty disgusting. I really don't like doing it and I am going to try to avoid ever doing it again.
Yesterday morning we were supposed to have a lab meeting, but the person who was going to be presenting hadn't finished her slides, so Calvin and I went to meet with Dr. Mao instead. He started by doing a basic overview of the purpose of his lab, and why he does both research and Clinical work for his job. Then he basically laid the table open and said that Calvin and I could either write a review, or do a mini-project of our own. If we wrote a review, basically what we would do is do a lot of reading and summarize something about the biology relating to what is being done in the lab. For example, we would do a bunch of research on the biology of a spinal chord, for example, because that would come in handy when trying to understand what all the images people were looking at of spinal chords meant with the differing expressions in proteins, etc, etc. But I'd really rather not just do a bunch of reading and then write a paper, because, firstly, I could do that anywhere, I don't need to be in a research lab, and secondly, it's the middle of the summer and I have written plenty of paper's this past year. Calvin was on the same page as me, so we decided we would figure out something we could do for a project. We went back to Dr. Mao a bunch of times with different ideas, and eventually we came up with a plan.
Our mini-project is going to be this: We want to know the effect on pain thresholds of rats when induced with depression by forced swim tests. This should take me to the end of July.
The problem left to me now is what to do with this week. I have 3 days left of work this week and nothing planned for me, that I know of. If they don't have anything for me to do I suppose I will research more into what the results of our experiment could mean, or I could always occupy myself by reading for my psych class.
Today, for example, I got here a bit late because I had to get a weekend slip from Harvard and the office doesn't open until 9am. Then I did a bit of research and wrote this post, and when I went to find Shuxiang, he said that he wasn't doing anything of interest today, just writing his paper, I think (again, it was hard to understand him). He can't continue with his research until a specific antibody comes in, which he has been waiting for for two weeks because it's on back-order. And they were done with the behavioral testing for the day, and won't do it again until Thursday, so I can't go watch it. So basically I am going to go eat lunch now, and then go back to Harvard, at 1pm. If I was a slacker and liked not having to do anything, this would be fabulous. But as it is, I'm just going to end up reading Harry Potter all afternoon, which I guess is fine, but I'm supposed to be at work... So I can't say I'm happy about it, really.
I get to shadow a doctor through the OR in MGH on Thursday, though! I'm really excited about that!!
Monday, June 22, 2009
Day one of Internship
I guess I wasn't so on top of things coming into this program, because when Wes and I got to MGH and got up to Dr. Dunn's office, he already had an ID and I was sitting there without my forms, which were supposed to be faxed in by two fridays ago. It wasn't entirely my fault, though. I hadn't been able to send them in because I was in England. But the lady at the desk had them there for me to fill out. However we spent about an hour and a half trying to get hold of people on the phone, attempting to get me an ID, and in the end I didn't even end up getting one. Now I need to find my way to the Charles River Plaza to get a TB test at some point, and until I get my ID, I won't be able to get into my lab without calling one of the people up to let me in.
There is one other intern there at the lab at the moment. His name is Calvin, and he's from Milton Academy. His dad works somewhere in the building I think, and that's how he got the internship. He told me to bring my laptop tomorrow. Apparently we get a lot of downtime.
The one thing I did today was watch three people from my lab (the only one whose name I remember is Connie) do an experiment in animal behavior. There were a bunch of rats, whose mechanical pain and thermoradiation responces they measured. Then, they do surgery on each of them (I don't know of what kind) and do the tests all over again, to see the change in pain thresholds. It was very interesting to watch, although since they were all talking in chinese, I attempted to learn how to do the experiment by watching instead of listening.
When I was eating lunch with Connie and another one of her friends from the lab, I saw Chris walking past me. I didn't know he was doing his internship in the Navy Yard too. Apparently he started last week and he said he did like hardly anything the first couple days, and then they started finding things for him to do, although he still has a lot of downtime, too. He also mentioned something about removing the bone marrow from a rat!
This internship is very free-form. Basically I can just shadow people and watch what they're doing the whole time if I'd like, or I can do a short project of my own, and just ask people if it's okay to use things. I won't be doing that quite yet, seeing as I don't really know how anything works, but Connie said that tomorrow she's going to be doing some immunohistochemical staining, which sounds cool, so I am going to look that up before tomorrow so I have a basis of understanding before she tries to explain it to me.
Hopefully tomorrow I have somewhat of a better idea what I'm working towards!
~Charlotte
There is one other intern there at the lab at the moment. His name is Calvin, and he's from Milton Academy. His dad works somewhere in the building I think, and that's how he got the internship. He told me to bring my laptop tomorrow. Apparently we get a lot of downtime.
The one thing I did today was watch three people from my lab (the only one whose name I remember is Connie) do an experiment in animal behavior. There were a bunch of rats, whose mechanical pain and thermoradiation responces they measured. Then, they do surgery on each of them (I don't know of what kind) and do the tests all over again, to see the change in pain thresholds. It was very interesting to watch, although since they were all talking in chinese, I attempted to learn how to do the experiment by watching instead of listening.
When I was eating lunch with Connie and another one of her friends from the lab, I saw Chris walking past me. I didn't know he was doing his internship in the Navy Yard too. Apparently he started last week and he said he did like hardly anything the first couple days, and then they started finding things for him to do, although he still has a lot of downtime, too. He also mentioned something about removing the bone marrow from a rat!
This internship is very free-form. Basically I can just shadow people and watch what they're doing the whole time if I'd like, or I can do a short project of my own, and just ask people if it's okay to use things. I won't be doing that quite yet, seeing as I don't really know how anything works, but Connie said that tomorrow she's going to be doing some immunohistochemical staining, which sounds cool, so I am going to look that up before tomorrow so I have a basis of understanding before she tries to explain it to me.
Hopefully tomorrow I have somewhat of a better idea what I'm working towards!
~Charlotte
Sunday, June 21, 2009
First Day
Hey all!
This is going to be my blog for my 2009 SFS summer adventures!
Just for general info, my name is Charlotte Garcia, and I will be doing an internship at MGH this summer, working with Dr. Jairen Mao in his research lab, which focuses it's research in pain management. I will also be living at Harvard for the 7 weeks duration of my internship, enrolling in a class there through the summer school.
I registered for Harvard Summer School yesterday. It was a really nice day, although there were really long lines everywhere we went. I went home last night and came back in the afternoon today.
I got to know my roommates a bit. There's Sarah from Texas, Min from California, and Olivia from Nevada. They all seem really nice! Olivia talks to everyone we walk past and is really outgoing. It's really funny.
So, I am going to my first day at the lab tomorrow! I'm really excited but also really nervous because I don't know quite what I am going to be doing there.
I'll update as soon as I know more and have the time! I think I have a pretty busy day tomorrow though.
~Charlotte
This is going to be my blog for my 2009 SFS summer adventures!
Just for general info, my name is Charlotte Garcia, and I will be doing an internship at MGH this summer, working with Dr. Jairen Mao in his research lab, which focuses it's research in pain management. I will also be living at Harvard for the 7 weeks duration of my internship, enrolling in a class there through the summer school.
I registered for Harvard Summer School yesterday. It was a really nice day, although there were really long lines everywhere we went. I went home last night and came back in the afternoon today.
I got to know my roommates a bit. There's Sarah from Texas, Min from California, and Olivia from Nevada. They all seem really nice! Olivia talks to everyone we walk past and is really outgoing. It's really funny.
So, I am going to my first day at the lab tomorrow! I'm really excited but also really nervous because I don't know quite what I am going to be doing there.
I'll update as soon as I know more and have the time! I think I have a pretty busy day tomorrow though.
~Charlotte
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