The End (Not Really)

My last post! Maybe. I have mixed feelings about this; this will be the last project I do for high school. I didn't think the day would come. It's weird to think that I won't be returning to BASIS, after going there since fifth grade. I suppose, though, you can take the person out of BASIS, but you can't take the BASIS out of the person.

I visited Tufts for a night and day. Even though I didn't really like the weather (though I suppose I'll have to get used to it. Cold weather actually exists - who knew?), the campus was nice. There was a great view of Boston from the library roof, and the dorms were quite spacious. And the food was good (for college food, anyways. I'll enjoy my home-cooked meals while I have them). I, however, always felt like I was walking uphill.

My grandma and I next to the new Jumbo statue. I think it makes me look even shorter.

You may be wondering why my blog post was late, and it's partly because of the Jumbo Day I went to. I also didn't want to post until I had made a final decision for my college. And now, six days before the deadline, Id like to say that, even though I'm not exactly tall, I'm proud to be a Jumbo! Going to the admitted students day really helped me become confident with my choice.

And now, drum roll please, what you've all been waiting for: my results!

I tested about six drug substances, with varying success.

Here's my data for AK11, the first drug that I tested:

Yay for pretty graphs.

The bars show the proton flux of the liposomes. You want the "blanks," the liposomes without M2 proteins, to have a low proton flux, because there should be no way for the protons to move into the liposomes when valinomycin is added. If there was proton flux, the liposomes would be "leaky" and I'd have to redo my whole entire experiment. You want the liposomes with M2 proteins to have a high proton flux, showing proton movement when valinomycin is added. And finallu, you want the liposomes with M2 proteins and drugs to have a lowered proton flux, showing that the drug blocked the protons from entering the M2 channels.

Although my bars on the graph look decent, my data was not the best. For one, I only did two runs of each type of liposome instead of three. (I know, I was lazy, but I only had one pH electrode. Can you blame me?) Also, the standard deviation bars, the vertical lines coming off the bars, are pretty big, showing that my data wasn't as consistent as it could have been. Finally, the proton flux is extremely low - partly because we weren't using the right concentration of M2 proteins to lipids, so the effectiveness of the experiment was compromised. But you live and you learn, right? (Except in this case, the learning took about a month, so I used to wrong concentrations for the first month I was there.)

I had one drug, AK40, that I didn't like that much. I couldn't make the drug dissolve that well, so some of my data was unexpected.

The graph style changed a bit. It was - surprisingly - updated. And - even more surprisingly - it worked better.

The standard deviation bars were even bigger on this graph, especially on the liposomes with drug. Since the drug easily precipitated out of solution, I think that sometimes I didn't add any drug into my liposome solution. So sometimes I got results for the M2 with drug that looked awfully similar to the results for just the M2. And, for anyone who is wondering why I just didn't repeat the experiment again, I'm just going to mention that this was my second attempt at testing this drug. (Ugh, I wasted like a week on just this drug alone.) So we decided that I should just test other drugs, and that maybe we'd come back to this drug.

If you thought that all my data was sub-par, then I'm sorry to disappoint you. (Not really.) I saved the best for last, although this graph will look a little bit different.

I'll give you a hint for the difference: the graph was not, in fact, "updated" again.

This graph actually shows two drugs that I tested. The first drug, AK13, was good at blocking the WT M2 channel, but only moderate at blocking the mutated S31N M2 channel. When I was testing, my mentor asked me to test the drug he had just purified, NAG107. Because I had extra S31N liposomes, I ran the experiment - and the block was pretty remarkable. (For clarification, the first six bars are the same as the earlier graphs. The last bar only shows the S31N M2 and the NAG107; I didn't rerun all the blanks.)

So, even though I didn't plan on testing the NAG107 in the beginning, I'm glad I did. I think that because it shows so much promise at blocking the proton flux, it's going to be tested on the oocytes. With any luck, it'll move on to animal testing as well. (And if it doesn't, well.... We have a lot more ideas for drugs. Some of them involve copper complexes. The AK drugs I tested were all amantadine derivatives.)

Overall, I got to see a bit of everything - good results, moderate results, results I wish I didn't get.... As my SRP journey draws to a close, I'm excited for a new journey to begin: college! I'm glad that I had the wonderful opportunity of working in the lab. I learned more than I thought I would. (Buses are pretty popular outside of Arizona, it seems like. Especially in Boston.) I think the lab is one of the few places that I can ask "why?" multiple times and not get annoyed looks. (Curiosity may have killed the cat, but the cat had nine lives.) Because of this experience, I want to get into research in college, and maybe after that. Life, it seems, is filled with uncertainties, and I just have to live it one day at a time.(Unfortunately, my time machine broke, so I can't live life backwards; I'm stuck living it forwards like everyone else.)

I hope you've enjoyed reading my blog as much as I've enjoyed writing it! (Is it bad that I've been highly amused by making songs into my blog post titles?)
- Lauren

She Said I Think I'll Go To Boston

T-minus two weeks, and counting. I wish I could have a moment of certainty and make a decision about the college I'll go to. But I suppose that would be too much to ask for. Instead, next week I'm going to attend an admitted students day at Tufts, so hopefully I'll closer to my decision soon. (But, let's be honest, May 1st is getting closer and closer whether I like it or not.)

Although it's been nice sleeping in, I miss going into the lab and doing something with my time. (Maybe I'll have to get a job soon so I have something to do. Yay...) All in all, I've had too much free time to sit around and think about the future. This includes the near future as well - it turns out I'm in the first SRP presenting group. I'll talk about my project and my experience on May 9th any time between 10am and 11am. At least I have something else to worry about now, besides college.

On a different note, my uncle took Dora to the vet this week. Since we're not the best people to take care of a wild dove, we gave her to the vet, who promised that she'd find a rehabilitation place for for her. It's nice to know that Dora will be returned to the wild, but she'll be missed all the same (except, probably, by my aunt).

I'm going to save my results for my final blog post (next week's post) and for my SRP presentation, so stay tuned. What will I talk about in this post then, you might ask. I asked myself this too, and after much discussion with myself (I'm not crazy; the voices in my head told me so) we decided to explain the graphs more. Before you get too excited (and yes, I can sense your enthusiasm), you should know that this means I'm guaranteed to add pictures along with the words, instead of just blocks of text.

That moment when it's easier to edit a picture in PowerPoint than Paint....

The pH on the side of the graph isn't exactly correct; the conversation from the voltage in the pH electrodes to the pH needs to be altered. The actual pH is around 6 to 7. We haven't fixed the conversion because we're looking for the change in pH, not the pH itself.

The first time I saw a graph like this, I was extremely confused. (And I didn't even have the arrows to help me.) I mean, sure, it looks cool - but it was just a bunch of peaks and valleys to me. If you're anything like me, you'd be wondering what, exactly, is this means in English. (And if you're not, I'm afraid you still have to put up with my questions and answers.)

Valinomycin is a potassium ion-selective transporter; it only moves potassium ions. It looks a little like this:

It looks so simple.... I've yet to figure out how it actually works.

The first graph I showed you is a blank, meaning it has no M2 protein channel, and it also has no drug added. The liposomes (hopefully) have more potassium ions than the external solution, while the external solution (in theory) has more protons than the liposomes. This sets up a gradient; the potassium ions want to leave the liposomes, and the protons want to enter the liposomes. However, without a channel, the ions can't move.

This is where valinomycin comes in. (Literally.) It's first added to depolarize the liposomes (meaning that protons enter the liposomes). It allows the ions to move - but only if there is a M2 channel. Therefore, in the blanks, we expect there to be no valinomycin signal (and by "signal," I mean change in pH). And, if you look back to the graph, you'll see that this is true. You could argue that there is, in fact, a small change in pH, but I'd tell you to look at the second addition of valinomycin. Both changes in pH are the same. We can assume that the pH change is from something else, instead of the uptake of protons into the liposomes. And it is; the valinomycin is in an ethanol solvent, which is basic. Adding ethanol to this experiment would produce a similar pH change.

Conversely, we would expect a valinomycin signal in a liposome with M2 channels.

Yay, the valinomycin signal is different. We're done now, right?

I'm afraid that there's more to the graphs than the valinomycin signal. (There're three different colored arrows; you should have expected this.) If you compare the graphs, you'll notice that not only did the valinomycin signal increase, the CCCP signal decreased.

CCCP is a proton-selective transporter, and it acts like a M2 channel. It allows protons to come into the liposomes, resulting in a small change in the pH of the external solution. In the blanks, we expect a CCCP signal, because the liposomes are not depolarized by the valinomycin. The CCCP depolarizes the liposomes in the blanks, instead of the valinomycin. In the liposomes with M2 channels, there is no CCCP signal, because the liposomes are already depolarized by valinomycin. (Again, the CCCP "signal" is due to the ethanol, instead of proton flux.)

The previous graph has liposomes with M2 channels, and the next graph has liposomes with M2 channels and a drug. Kudos to the person who can guess what kind of "signals" we're looking for. (Hint: A good drug will block proton flux.) (Another hint: CCCP will depolarize the liposomes, whether or not there are M2 channels present.)

Am I the only one who would look at the graphs and this painstakingly detailed explanation, and ask what the HCl is for?

If you guessed that a good drug would reduce the valinomycin signal, you win! (If you didn't, at least you tried, right?) My whole project was based on the comparison of these graphs. I put my data into an Excel spreadsheet, and somehow they got an estimate for proton flux during the addition of valinomycin. Unfortunately, the numbers were pretty subjective, since we changed the numbers for the graphs ourselves by moving the red lines near the valinomycin signal. Fortunately, someone in the lab is working on a program that will automatically take the differences in the pH before and after the addition of valinomycin into account, so the numbers will be less subjective. Naturally, this program will only be used after I've completed my project. (I suppose that's the way the cookie crumbles.)

 

Thank you to anyone who even attempted to read this! I appreciate your efforts. (And if you just skipped to the end, and saw the word "cookie" - well, no cookies for you, then.) Feel free to ask me any questions.

Until next time!

- Lauren

It's the Eye of the Tiger, It's the Thrill of the Sight

A few weeks a go, whenever someone asked me where I wanted to go to college, I replied, "I don't know." I didn't want to make a decision until I knew everything - whether I was accepted, what the financial aid was, etc.. I wanted to make a fully informed, logical decision, so I didn't let myself get too attached to any one college. However, I'm not entirely sure this was a good idea (although, if I could redo everything, I'd do the same thing), because now if you asked me where I wanted to spend the next four years of my life, I'd still say, "I don't know." I've discovered that I'm a pretty indecisive person (at least I learned something from this process). I have 20 days to make a decision (but who's counting?), yet I hope I'll have a decision next week.

Even though I've now spent almost six weeks in Utah, I still managed to learn new life lessons this week. My aunt went away this week, and she left some meals for me to bake for dinner, so I learned how to not burn food (Yay me). Four meals down, one meal to go. I also learned that there's something more awkward than walking through a crowded college campus as a high school student: walking through an empty college campus as a high school student. It turns out that UVU, the campus I walk through to get back to the house, was on spring break this week, and I didn't realize it until I had walked halfway through the building. (It's amazing how self-conscious you can become even when there aren't any people around.) But the doors were open, so I don't think I was doing anything wrong. Finally, I learned that I really shouldn't be surprised when random things happen. For instance, I woke up on Tuesday to be greeted by this sight:

Say hello to my new pet dove, Dora.

My uncle had apparently found her on his morning walk. Her wing was injured, and she flew right in front of him. She even let my uncle carry her back to the house and put her in this cat carrier. We took her to the vet on Wednesday, and we're trying to heal her wing, which looks like it was scratched by a cat. We're not sure if she's domestic or wild, but I think, if her wing heals, we'll put her back outside. (And by "we," I mean my uncle, since I'm leaving on Sunday. Sadly. I've become to primary caretaker of her.) She seems fine besides her wing, but I don't think she's eating much. Hopefully she'll be okay.

This week was bittersweet, since it's my last week in the lab (but, (un)luckily for you, I still have a few more weeks of blog posts, so stay tuned). Fortunately, I finally learned the liposome-creation-and-testing process well enough so I can test three to four drugs per week, if not more. Unfortunately, it's a bit late to apply that knowledge, but better late than never, I suppose. Looking back at my first month here, my results just seem so slow.  Half of the results I've obtained have come from this week alone; the other half come from my previous five weeks here. On the bright side, I'm glad that none of my tests went awry yesterday. (I was told that apparently everything goes wrong when you're leaving soon. It's like the liposomes know you're leaving.... My gloves, at least, knew I was leaving; I used the last of my extra-small gloves today.)

This week, I  had - not one, not two - but three pH electrodes to work with. ( I never thought this day would come.) It was, needless to say, very exciting to finally test liposomes in sets of three, instead of one test at a time. (This is also probably why I was able to test so many more possible drugs. Maybe if they had fixed the pH electrodes sooner, and I would've tested more possible drugs.) I'm sure you're all wondering how pH electrodes can be "fixed," so here is our wonderfully high-tech solution (as usual) to the pH electrode noise/artifact problem.

Yes, that's tinfoil on the pH electrodes. Welcome to the 21st century.

Something with the metal-to-metal contact must have worked, because all three pH electrodes now have no noise, and very minimal artifacts. I'm not entirely sure this is a good permanent solution, though. The tinfoil seems to easily break and sever the metal contact.

On another note, I have found the one working new pieces of equipment that we have in the lab (besides, of course, the building itself). I'm not entirely sure how expensive it was, but I think it was a few grand, so I'm glad it actually works (though it took a few weeks to be calibrated).

And you thought your dishwasher was expensive.

I was going to talk about my most recent results, but I think this post is already a decent size (wow, the words go by fast). So, you're welcome; I won't confused you any more (for now...).

Until next time!
- Lauren

Why You Gotta Be a Fool? Don't You Know I'm Human Too

Wow, a lot has happened this week. Although I got rejected from my "reach" schools, I still have quite a few options for colleges. I'm having a tough time deciding what to do. I wish I could see into the future, but I suppose life is more fun when it's surprising.

Anyways, it's amazing how much can change in a few weeks - and how much stays the same. I'm glad to be back in the lab, actually doing something. (As an added bonus, it keeps my mind from over-stressing about college decisions.) But, early Monday morning, still groggy from getting up at 4am, I walked into the lab hallway to be greeted with a sign. You know it's a Monday when you see this:

How, exactly, you can close a hallway, I'll never know.

I didn't want to walk through that hallway, anyways. Thanks for asking.

I tried to make more liposomes on Monday, but in hindsight, I should've just waited. I spent at least an hour preparing my lipids and M2 proteins, and everything was going fine. I was even told that we got more internal buffer (it was running out before I left. I actually thought we weren't going to have any when I returned). Unfortunately, somebody didn't put the red tape back onto the internal buffer.

So I may or may not have put 1M NaOH into my lipids instead of internal buffer. Whoops. (I'm sorry, liposomes. I failed you.) How can such a small change of tape color make such a difference.... (I know, I know, I should've read the label on the bottle, but I wasn't expecting it to change.) At least this mistake shows that I am, indeed, human.

I was also told that we had three working pH meters now. Sadly, I ended up doing my tests - or at least trying to do them - on April Fool's day. I feel like the world wanted to prank me, so, naturally, all three pH meters weren't working. I mean, I know computers don't like me, but can't pH meters at least tolerate me? (Is that too much to ask for?) So Wednesday was spent trying to make them less noisy and less drifty, with moderate success.

On the bright side, some of the pranks I saw were highly amusing. Especially this one.

Notice how there are posters and even a lamp.

What are the handrails doing in an office, you might ask. This "office" is actually an elevator, and the teacher spent his day greeting unsuspecting students as if they had all come to his office hours. (I suppose that's one way to get a higher turn-up for office hours....)

On Thursday, only one pH meter was working (I really shouldn't be surprised), which meant it was a long day. One of the pH meters had almost no noise, and I was excited to use it. It wasn't meant to be, however. I discovered that if I leaned on the table, the two new pH meters would exhibit "artifacts," basically spikes in the pH readings that are not from pH changes in the solution. I thought I was onto something important, until my uncle informed me that this effect was common in pH meters.

Basically, my body was acting as magnetic field, which interfered with the voltage readings from the pH meters. To solve this, we have to ground ourselves, or the pH meters. (One solution involved me wearing a metal bracelet, which was strapped to some kind of "ground." I felt like I was chained to my experiment....) I don't think we've found a permanent solution yet, but I hope that we're on our way. It's odd that only the new pH meters are affected by my proximity. (Such is new technology, I suppose.)

Speaking of new technology, when I was gone, my lab decided to update the Windows 2000 computer, which is the computer that records all the pH changes. This lab must be allergic to new technology or something, because apparently the new program crashed, and brought down the program I used with it. Thankfully, we had another similar - and older - program that I now use.

Don't worry, though - this means that the Computer Says game just became more interesting! Only now, the correct information isn't even displayed on the computer; I had it written in my lab notebook somewhere.... The computer beeps one sometimes, usually before it thinks I'm supposed to do something, then kindly beeps three times ten seconds later. I think I'm becoming more adept at ignoring the beeps. (And this is coming from someone who rushes to get everything out of the fridge before the fridge beeps as a reminder that it's been left open.)

In other news, my results from testing AK40 again were inconclusive. Sometimes, it seemed like I had drug, and sometimes the drug was suspiciously absent. I suppose I should've seen this one coming; the drug was hard to get into solution. My mentor thinks that there was a very small amount of drug precipitate that wasn't in solution. So, only one thing to do from here: make more liposomes.

On the bright side, at least my graph from AK11 (the first drug I tested) still looks nice.

Until next time!
- Lauren

All The (Incredibly) Small Things

Last week felt oddly empty. Maybe it was because I wasn't in the lab, or that I wasn't constantly thinking of (hopefully) funny things to put into my blog. (Or maybe, I actually missed blogging in general. What a weird concept to think about.) Either way, my break was relaxing. Mentally, at least. It was pretty physically challenging with soccer games, so I suppose these balance each other out.

On a separate - but not any less important - topic, the decisions are coming, the decisions are coming! I'd like to thank all the "reach" schools I applied to for waiting until the last moment to release their decisions (some will be released as late as March 31st in the evening). I guess this is all a lesson in patience. 

Since not much new has happened, I've decided to go back to the basics for this blog post and describe how the influenza virus works. I don't claim to be an expert in this subject, so bear with me.

Sadly, I didn't draw this. (Surprising, right?)
 Credit to: http://www.nature.com/nbt/journal/v28/n3/fig_tab/nbt0310-239_F1.html

In replication, the virus first binds to a sialic acid-contating receptor on the host cell, which triggers the endocytosis of the virus. On a side note, do you recall that the types of the flu are labeled as HXNY, where X and Y are numbers? (If you didn't, now you do.) The "H" part of this - the hemagglutinin (HA) part - is responsible for the binding of the virus onto the host cell. There are around 18 known variations of the HA, although only a few, such as H 1, 2, and 3, are common in humans.

Once the virus is engulfed, it's inside an endosome that has a pH of about 5 or 6. Because of this low pH, the M2 channels in the virus allow protons to enter the virus, which then changes the virus's conformation, essentially activating it. This is the step that my lab is trying to prevent. Block the M2 channels, block the proton uptake - and therefore block viral replication. (In theory, anyways.)

The pH also allows the virus to release its eight single-stranded "negative-sense" RNA into the cytoplasm and, eventually, nucleus. In this stage, new HAs can be formed when two different strands of the influenza virus, such as a human and avian strand, infect the same host. The RNAs are mixed, and the viruses that are produced contain RNA from both sources. The "negative-sense" means that the RNA cannot be directly made into proteins; first, it must be translated into "positive-sense" RNA. (At least this naming makes sense.)

The host cell makes more "negative-sense" viral RNA. The virus even prevents the host cell from making its regular mRNA, so the host cell is basically converted into a virus-producing factory for the rest of its short lifespan. After enough viruses are produced, the cell breaks down. Poor cell.

Viruses are produced when they "bud" off of the host cell. In doing so, the RNA is enclosed in the host cell's membrane, along with the HAs and neuraminidases (NAs) - the "N" part of HXNY - that the host cell also produced. (You totally remembered these too, right?) After the "budding," the virus is still attached to the cell through the sialic acid. The NA cleaves the sialic acid, releasing the virus, which can then go on to infect more cells. (More viruses. Yay.)

And that's the process. Simple enough, right? If not, I'm always open for questions. In the meantime, good luck to anyone who's waiting for more college decisions!

Until next time!

- Lauren

Shake It Out

These last few weeks have been filled with learning experiences. I think one of the biggest lessons I've learned is that the bus waits for no one. Although this lesson was a hard pill to swallow, I've (mostly) accepted my fate of missing the bus. Fortunately for me, the bus comes every 15 minutes. This, however, doesn't make me feel any better when I'm about one minute from the bus stop and the bus suddenly pulls into the bus stop, waits ten seconds (if I'm lucky), then continues on its path.

Also, I've taken the lack of comments from my blog group on my last post to mean that they don't mind if my blog posts get to be a bit longer, or if I decide to start posting more often. Thanks, guys!

As for new experiences this week (since missing the bus isn't exactly new):

My cousin and I in bubble balls.

Enough said on that topic.

Earlier, I was able to watch tests on oocytes (frog eggs). Since I'm not a student at BYU, I can't really work with living cells, but I'm still able to observe (and, of course, ask questions). My trainer, Kelly, injected oocytes with the full length M2 protein, then waited a few days for the cells to express the channels. His results will be a bit more accepted than mine, mostly because oocyte tests are common, while make-your-own-liposome tests are few and far in between. Anyways, moving on. When he was ready to test his oocytes, I followed him into the lab next door and saw this.

How come their death ray machine is better than our death ray machine? We have to step up our game.

I'm glad that I was able to see this experiment, though. Apparently Kelly has been working on this setup for months, and he's only started getting decent results recently. (And, as a side note, I think that we're eventually getting one of these in our lab. Yay for two death ray machines.)

After much more discussion, we've decided to change the M2 protein and lipid concentrations - again. I hope this is the last time, but I wouldn't be surprised if it was changed again while I'm gone for two weeks. (I'm going back to Arizona for some prior commitments, but I'll come back to the lab. And yes, in case you were actually worried, I will still be making blog posts) Naturally, we didn't decide on this until after I made my liposomes, so I'll make liposomes with the correct concentrations when I return.

Everything mostly ran smoothly this week, so I can't really complain. I learned, however, that chemicals and equipment tend to run out rather quickly, especially when you're not in the lab. So I spent some time on Thursday making new CCCP and valinomycin solutions, and I'm willing to bet that those will be gone upon my return. Such is life in the lab. I also learned that some substances just don't like to be dissolved. It turns out that most of the drugs I'm testing have hydrophobic groups.

This week, I think I've almost perfect my vial drying method. Since any water droplet in a test tube or vial could dilute our solutions, we have to shake them out and let them air dry until all visible water droplets are gone. The shaking includes vigorous amounts of arm shaking, along with a bit of wrist flicking. And the best part is, I haven't accidentally let go of the vial while drying it. (Yet.) (It's amazing how much our technology our methods use. It's almost as if I'm back at school.)

Because seven out of my eight minutes of actually testing liposomes is filled by me staring at the clock, waiting for the right time, I've come up with multiple ways to keep myself entertained. Sometimes, I'll try to clean up as much as I possibly can (or add pipette tips to the box) before my minute is up and I have to add something to the liposomes. Other times, it's as if I'm playing Simon Says, except it's Computer Says. I have to make sure I follow what's on the computer screen. To make things more difficult, the computer beeps at different intervals (it was used for similar testing, but the process has changed, and no one bothered to make the beeps on time). I think I'm winning this game of Computer Says so far.

Four out of six of my liposome samples this week were great. As for the other two.... The results from the S31N without drug and S31N with drug look exceedingly similar. Which isn't really that good. The drug is either horrible at blocking this mutant M2 channel, or there's something about the higher concentration of M2 proteins and lipids that's not ideal. Either way, I'm sure I'm going to be testing this same drug when I get back, just to see what the problem is.

I'm sure I'll have plenty of time to explain what "good" and "bad" results are later, but for now, I'll explain about pH meter "noise" (I'm sure I could explain both, but this post is getting to be longer than expected. Hopefully no one minds).

With "noise."

Without "noise."

Pictures, I think, speak louder than words. And maybe you can now appreciate the small scale that we're working on. Also, the numbers on the left are not the actual pH; they're a voltage of some sorts. The pH is somewhere in the 6 to 7 range, but the change in the voltage and the change in the pH are the same.

And happy early pi day! I wish I had been born tomorrow at 9:26 am.

Until next time!
- Lauren

P. S. I think I'm going to claim next week as my "spring break" since I'll be playing soccer all of next weekend, so if I don't post, that's why (I know, I know, I can sense your disappointment).

It's a Tall World After All

I learned earlier this week that my brother, sister, and mom are getting their motorcycle licenses. I leave for three weeks, and this is what happens.... I hope this means I'll get the car though. I could live with that.

On another note, it actually snowed this week. I can't even remember the last time I woke up and the ground was white. I never thought I'd say this, but I'm glad that it's back in the 40s again. Who knew that I could actually enjoy weather below 80 degrees (what is this blasphemy?).

As for new experiences, this weekend will be interesting. I can't remember the last time I had to change my clocks for daylight savings. What kind of nonsense is this? It just seems so inconvenient to change the time and lose an hour of sleep. It's not like the sun is staying in the sky for an extra hour or anything. I'm glad Arizona doesn't follow daylight savings - but then again, we get enough sun as it is. Why would we inconvenience ourselves for "more"? (Okay, maybe I'm a bit mad that I'm losing an hour of my day.)

I think my body has an internalized alarm clock, but I'm not sure yet. I'm not used to getting up by myself in the mornings since my mom usually wakes me up. (I know, I'm 18 years old, and my mom woke me up for school. And while I'm at it, I'll say that my dad still made my lunch. Sue me.). Whenever I set my alarm, I think I subconsciously get anxious that I'll miss the alarm, so I wake up around 30 minutes before it actually goes off. Does this happen to anyone else?

If there's one lesson I learned this week, though, it's that labs are not meant for short and/or small people (even if they're meant for working with small measurements). I mean, the pipettes can measure under 5 μL of fluid. I've never seen beakers so tiny either.

Aren't they so cute? Shh, don't tell Kelly (the person training me) that I said that. He thinks it's weird that people call beakers cute.

There are even smaller flasks than that. The two pieces on the left measure 25 mL. I couldn't find any dirty 10 mL ones to borrow for a picture. At least the lab has extra small gloves (but those are still too big. There's no winning. Sigh.). You would think that with all of these small things, that maybe there's be an extra small lab coat. But I guess that's too much to ask for. Don't worry, though; they still give some consideration to short people.

See? I get my own stool. It was just kind of  lonely so I claimed it.

Okay, I think I'm done with my (not so) passive aggressive ranting. Maybe. But I have good news! All my liposomes were good this week, and I had to make six vials instead of four. I'm two for two so far. I wasn't so sure about my S31N liposomes (which have mutated M2 channels that prevent amantadine from binding) because I left the dried lipids out for a few hours. I wasn't expecting to do that, but when I couldn't find any S31N protein, I had to ask for more. Which led to the S31N samples being put into the photospectrometer. Then I had to learn how to convert the absorbance into an actual concentration. I suppose I wouldn't learn as much if everything was easy.

On this note, I have more good news (surprising, right?): we received new pH meters on Wednesday.

This is where I test the liposomes. The computer is good for something, it turns out. It tells me when to add chemicals, and then beeps at the wrong time intervals. As long as I just look at the information and ignore the beeping, I'm fine.

However, I learned that new does not necessarily mean improved. One of the pH meters was too "noisy," meaning that there was too much fluctuation in its readings. And the other meter just didn't work. So I only had one pH meter to use. To test six different vials. Twice each. And the tests last about 10 minutes every time. Yay. When I think about it, the situation is kind of funny. We get new technology, and it doesn't work, but the older technology works just fine. Oh, the irony. (Maybe I should use floppy disks and Windows 2000 more often.)

Overall, I think this week was filled with too much waiting. I had to wait for the concentrations of the M2 proteins before I could resuspend my lipids in solution. I had to wait to see which concentrations of the lipids and proteins that I should use. I had to wait for the filter supports to arrive (which they did on Wednesday). (Insert more discussions on lipid and protein concentrations here. I think we're back to using what I originally had in my last post. That's what I used anyways.) I had to wait for more CCCP because I ran out in the middle of my experiments. Thankfully, we found some in the freezer. (Not before I put everything away, though.) But, since my liposomes ended up okay, I don't think I can complain much. I just hope that I can test a drug that'll be a decent block against the mutated viral proteins.

I always feel like I have so much to say (well, to write). Maybe I should start posting twice a week.... I'm not sure my blog group would be too happy with that. Oh, well.

Until next time!
- Lauren

P.S. Success! I still managed to finish this on Friday, even if it is late. I hope I can keep my goal to post on Fridays to be somewhat consistent.