The little moments that make science (or anything) worth it.

     So right now I'm in a particularly good mood.  The reason for this?  I figured out something that had been plaguing me for a good month now.  Basically we are trying to look at our bacteria using a special microscope after tagging them to see if they have more or less of a certain protein on their surfce after mutating it. Essentially we look at the bacteria and more glow = more protein, less glow = less protein (simple right).  The only problem was we couldn't find our bacteria using this special microscope.  We did a roughly 12 hour day only to find at the end that we couldn't see our bacteria...frustrating.

      Fast forward over a period where I put off working with it (cause I'm a bit over it) and am trying to get permission to use another scope we used in the past and I know works.  We decide once we know we can't use the old scope to try again only we won't do the whole 12 hour process, we just want to see if we can find the darn bacteria.  So we use a normal microscope and find them in a minute or less.  So far so good.  We then go to the special microscope (which is inverted to allow the use of larger sample containers etc) and try all sorts of different things but still can't see our bacteria.... more frustrating.

     That is until we accidently move it to a larger objective and find them quickly.  Weird but encouraging.  We still need to get the high power objective working so that night I download information about the objective and scope.  The objective has a little marking on it (among several others): WD 0.17.  As I am finding out information about the objective I find out that WD stands for working depth of 0.17 mm.  Hmm.... interesting.  I mull that over in my head over night and suddenly it hits me.  Working depth... that's as far from the objective that it can see.  This is an inverted scope so we were trying to have the objective look through the slide which is thicker than 0.17 mm.  I bet all we have to do is invert the slide and we'll see our bacteria.  Today I invert my slide and what do you know?  I see my bacteria!  Hurrah! One problem down, many more unknown problems ahead.  When we were trained on the scope we were taught by people who only need to use the larger objectives with much longer WD values.  Lesson, it pays to keep thinking about things.  (Cartoon bellow).

     This type of eureka! moment has happened one other time in my time here.  I was trying to generate double mutants but kept on getting single mutants.  After much thinking I finally figured out that the mutations were too close together and I would overwrite one when trying to make the other (cartoon bellow).  These small realizations are what keep me coming back.  There is something about these realizations that, while they seem obvious after you have figured it out, make all the blood sweat and tears worth it. :)