Monday problems

Check on cells or make coffee.
Check on cells or make coffee.

Sit paralyzed at desk wondering whether to check cells or make coffee?

Yep, it's Monday.

That awkward line between tech and student

The day has come. You're all set up to do an experiment. You've spent days preparing. All of your pipets have been lined up the night before. Tip boxes are full, waiting. Media is warm, gloves are out...and your PI walks by your bench and demands that you attend a seminar with him (or her.)

(Alternate universe: Your PI emails you the week before alerting you of the talk and you're really thankful because you didn't have that much to do that day anyway.)

EITHER WAY--you attend dutifully.

On the way to the seminar (in which you lead the way because Techs Know Everything including all the shortcuts between buildings), your PI reveals to you that the seminar speaker is a moron, has submitted a moronic paper to a moronic high profile journal, in which your PI is in the middle of reviewing. But of course he doesn't want to affect your view (yeah right) but instead wants your thoughts on the talk afterwards. (No pressure!)

Tech-PI relationships are a funny beast. It's a fine line between employer-employee (which it is at face value) and mentor-mentee (which it may evolve into). Each has it's own unique sets of issues. I'm in the lucky situation where my PI knows that this job is just a step in the direction of a career for me. He knows that I'm probably going to be leaving soon to go off to grad school in...something, or try to move up and out into a new position...sometime. When your PI knows that, and better yet, respects that--that's when you can start pushing the boundaries of employer-employee into mentor-mentee territory.

However, a big thing to keep in mind is: Cave Canem...ma.

source

Or, for those not well versed in Latin/English (Lat-glish?), beware of dogma.

I think a lot of scientific disagreements, the higher up you get on the ladder, tend to be fueled by competition and ego more than anything, while the differences are more or less just differences in dogma. But vastly different ways to look at problems are more or less what fuels scientific discoveries. I mean, someone has to be thinking outside of the box. I guess if you're the tech you just hope it's your PI.

It's a funny thing...and I'm having trouble writing it now because I'm trying to say it as delicately as possible, but keeping an open mind is the most important thing you can do in this situation. (Apart from staying awake during the actual seminar. Obviously that is priority numero uno.) At this very very very early stage in my career, the most important thing I can be doing is looking for places to think critically. Both of my PI and of his competitors. It's not always easy, and it's obviously very important to keep in mind who's grant you're being paid off of, but it's also really important to remember that thinking critically about something is not synonymous with criticizing it. And dogma and broad generalizations and effective public speakers get funded, because sometimes making things seem black and white is the best way to make sure your research gets noticed...but that's not I as a tech or a student, or whatever weird limbo position I'm in now as I'm starting to have my own ideas and taking ownership over my own research...that's not necessarily the position I want to be stuck in.

I wonder how other people in my position deal with this kind of thing. Thoughts?

Adventures in photobleaching

There are a couple of ways to act when you make a Really Big Mistake in lab.

1) Correct it, tell no one, and pretend it didn't happen. This method is preferable when you do something small and stupid. Like one time some sequencing results didn't look very good. And I realized that I had completely diluted my primers to the wrong concentration. Like, mMols off. This would be an okay mistake not to tell anyone else around you in lab, just as long as you fix it and make sure never to make that mistake again. (And also that no one else will be using your primer stocks.) (This also isn't exactly how it went for me because it took someone else pointing out that I had made a blaring error in my primer dilutions, so everyone knew about it in the end.)

2) Admit you made a mistake, ask for help, correct it, and then pretend it didn't happen (but never make that mistake again.) This method is especially useful when something big is going wrong, and you can't figure it out on your own. This is when you just have to suck it up that you'll look like an idiot for about 5 minutes while someone is explaining what you did wrong (or as I prefer to see it, what you weren't doing right!) but in the end it's all for the best because a) hopefully you learned something, and b) you won't make that mistake ever again. And it's a win-win! Because when you come across another poor soul who's made the same mistake as you, after you help them figure out what they weren't doing right, you can choose whether or not to admit to them that you too made the same mistake.

3) Ask for help, figure out that you were a complete idiot about something, and laugh uproariously about it. This might include just yelling PHOTOBLEACHING every 20 minutes or so for the rest of the evening after you figured your problem out. 


So this may or may not have happened to me today.


I will start off by saying that my lab does not do fluorescent microscopy. No one in my lab has fluorescent microscopy experience, and so we're looking at some fluorescence in our cells (just to check transfection levels) and using a scope in a lab downstairs that we've never really been formally trained on. The one caveat is that I come from a family of fluorescent microscopists--so this what follows is sort of unforgivable.

Here is what happened.

We walked downstairs with our little flasks of cells, and we put them under the microscope. We turned on the light, turned on the laser, made sure we were on the right filter, opened the shutter, and looked at our cells.

"Ooh!" My post doc says, "Pretty glowing cells!"

"Awesome," I say, "Let's put it on the screen!"

(I am clearly paraphrasing here.)

So, with the fluorescence still on, we manually switch the scope from eye to camera, we press "Live", and we wait for the picture to come up on the screen.

(If you're keeping tally of time blasting cells with fluorescence, we're now clocking in at 2 minutes.)

It takes another few seconds for the image to come up, then of course we have to refocus slightly for the image to be in focus for the camera.

Then we take a picture.

(Fluorescence still open, in case you were wondering.)

Then we hmmm and hawww and write down the settings we used so we use the same settings for all the other flasks.

Then we go get another flask.

I will skip the rest of the nitty gritty details, because the short of it is, we kept looking from eye to camera and back again, and since apparently neither my post doc nor I were feeling incredibly competent today, we kept forgetting to manually change the scope from eye to camera, all with the fluorescence on. 


And then we wondered aloud to each other, "Why are all of these flasks getting progressively dimmer?" And of course, being scientists, we say, let's get that first flask out that was really bright!

So we get the first flask out and everything is really dim.

In retrospect:

It didn't even occur to me that this was the problem until later I was talking about what I was doing to someone who actually knows how to do microscopy, and they were just like, You focused with the fluorescence ON?? My post doc still isn't quite convinced that this is the problem, but I think she just doesn't want to admit that it's the problem because it makes us look like complete idiots.

And me?

I am perfectly ok with looking like a complete idiot. Because, you know, how else would you learn? (By actually listening carefully to what people tell you to do so you never make mistakes? Where's the fun in that?)

And for the rest of the evening, yes, I have yelled PHOTOBLEACHING aloud at odd intervals. Because, arrrghhh.....PHOTOBLEACHING!