Tuesday, November 24, 2009

You know your experiment may in fact be too big when...

1. It takes nearly an hour to simply label all the tubes you will need for your experiment.

2. Once your raw samples are in their first set of tubes, they take up three ice buckets (two is typically a large experiment).

3. Your processed samples take up five sample racks (I think the most I ever did before was three).

4. You work for 6 hours during the day, go study for an hour and a half, take a test, go home, shower, come back to work to then work another 5 hours. Being in lab at 0330 is a wee bit creepy.

5. You still have two hours of prep work the next day before you can even think about running your samples.

Note to self, never ever do a take down with 30 mice, 60 organs, and 180 processed sample tubes again!

Tuesday, November 10, 2009

It only took four and a half years...

But I finally worked my first code. I know, ridiculously long time for being an EMT and not having worked a code, but then again, I have always worked low call volume places. It was an interesting mix of emotions, to say the least, and surprisingly ended on a good note.

Tones went out at the crack of dawn, and I was uncharacteristically awake and rested the moment I sat up (I am NOT a morning person in the least bit and have a very bad habit of staying up late when I am at the station). Ambulance (first!) and engine go out with only a minor hitch and I know the medic unit will be responding.

Get on scene, trying to orient myself as well as possible (I am the "Aide" on the ambulance) as our entire station basically moves itself into the house. We find the patient in the back bedroom, relatives trying to do CPR, frightened as they say they had checked on her an hour ago and she was fine. Something had happened in that hour as we find the woman, mouth open, lying bed with no movement. Warm. No breath. No pulse. No cardiac history. In fact, very little wrong with her to begin with.

We get her on a backboard and get to work. I end up with the bag valve mask (BVM) trying to ventilate her as one of our firefighters starts CPR. I check the airway, clear, and try to ventilate. I can't get the breaths to go in, adjust the airway and add an oral airway. Still nothing. Great.

The characteristic 'Stand Clear' of the AED rings in and my partner snags my shoulder to pull me back. I can't tell you enough how much I love my duty crew, because we always have each others backs. "No shock advised." Well that's odd, but we get back to work, still no air going in, as we make it to the stretcher outside and into the rain. Did I mention it was raining and cold too?

Still ventilating, we hand her off to the now present Medic Unit and they go into full swing. I can feel myself still jazzed on adrenaline as I look around and suddenly do a double take. One of the firefighters from my station and the current shift is present who was not at the station last night. Did I miss something??? Later find out he was on one of the other engines that was called to the scene.

Seeing as the Medic Unit had our stretcher and some of our staff, we follow them to the hospital, and I hate to say it but I fully expected our patient to be declared dead at the hospital. While I am a very optimistic person, I just didn't think this was going to end well. After a few moment of getting things together in the E.R., I poke my head into the room they wheeled her into and see the last thing I had expected. A beautiful EKG with a pulse of 74. I just blink. We saved her? I thought that almost never happens?

The Medic said that once they intubated a pulse came back, which makes me wonder if she had a pulse and we missed it. It would be a good reason why the AED wouldn't shock. I have so many more questions than answers when it comes to what went on in the back of the Medic Unit but I don't know if they will be answered any time soon.

During the entire event I felt oddly calm but wired at the same time. I know I came across as a bit frazzled but, honestly, it was the first time and I was trying to rapidly go through everything in my head. I know I could have done better. I know I should have done some things differently, but in the end, she didn't die and that is what counts, though I think I will always remember how we found her in the room.

On a lighter note, I was trying to get a replacement oral airway from the E.R. staff (we restock at the hospital) only to have this conversation.

Me: I need a purple OPA.

Nurse: Purple? What size is that?

Me: I don't know, ::shrugs:: purple.

Nurse: Hrmmm, does it have a size label on it?

I go grab one from one of the numerous bags we have to check.

Me: Nope, its just... purple.

In the end, we matched up sizes and got the right one, but its one of the many moments I love. Our OPA's are color coded to be idiot proof. :)

Sunday, November 1, 2009

Western Blot Woes (And Happy Endings!)

Not an EMT post to start, but at least this lab one has a happy ending!

To preface this post, one of the fellows in a neighboring lab told me this when I was trying to work this assay out months ago. “For a technique that is so commonly used in biology to prove ideas, it is by far the most fickle technique used.”

Day 1: Extract protein, quantify it, prep it, load it on a gel, get it to a membrane, incubate it with antibody to find protein of interest. A day of far too many samples with so little reward. I can’t even tell if I have screwed anything up yet!

Day 2: Finalize antibody stain to visualize protein with fun florescent molecules. After a bit of trial and error, produce a good looking film (yup, its just like x-ray film, but without the x-ray part. Photo sensitive instead).

But there is a problem. The antibody for my protein has previously, and consistently, been giving me only one band when I do this technique. Now I have two bands, at least about the same size (which is key), but two bands nonetheless. The difference between this run’s source of protein and the previous run’s source of protein: minimal.

I haven’t the foggiest why I now have two bands at the right size.

Day 3: Repeat staining process with a control protein antibody. Prep and visualize. Perfect film. Well, at least I know my technique isn’t off!

Day 4: Quantification. After imaging films, followed by far too much thought, and a fair amount of math, pretty graphs get to be formed! And on top of the fact that graphs in and of themselves are pretty, IT DID SOMETHING! My assay, done months ago with the cells saved in a -80°C freezer and a week spent working on the same Western Blot, did something!

I cannot begin to tell you how happy this makes me! Particularly since this means we may now have a mechanism for one of our models, and having a mechanism can be just the thing that is needed to get a paper published!