Lab Queries: What to Do When Things Go Wrong

I’m sure you’ll all be happy to know that this week on the blog we are leaving calibration curves and math behind us!

Don’t worry, if you still have questions about instrument calibrations, or a new issue arises in your lab, I’m always here to help you address those.  In talking to some of you at FELC a few weeks ago, the issue of troubleshooting came up a few times.  I know this can be a hard topic and really digging into a broken instrument can be an intimidating idea for some people.  But I’m going to let you in on a secret….it’s maybe one of my favorite activities to do!

I’m weird….I know…..I’ve accepted it and moved on. 😊

What to Do When Things Go Wrong

We’ve all had that moment….we started samples on an instrument, left for the night expecting to come back in the morning to a nice, completed data set, and return to find that our instrument malfunctioned!  Now results are behind schedule and you have the added issue of an instrument that just refuses to cooperate.

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Troubleshooting can be a frustrating and tricky activity to complete, especially because instruments never break at convenient times.  If I had a dollar for every time an instrument broke when I had rush samples to do, I wouldn’t have to work anymore!  But, because instruments will always continue to break, and I need to keep my job, I thought I’d fill you in on some of my favorite troubleshooting tips.

  • Follow the Path

A good way to make sure you don’t overlook any parts of your instrument is to start at one end of the instrument and follow the sample path through to the other end.  It’s easy to jump from place to place first, maybe checking your sample loop before jumping up to check your eluent filters.  Odds are that you’ll over look something, and your issue will be in the place you over look.  Following the path helps ensure that you give every section of you instrument it’s due investigation.

  • The Tubing Checker

One of my new favorite troubleshooting tools is a spare autosampler syringe!  When I need to troubleshoot one of my instruments with a large amount of tubing, I’ve found that using the syringe to carefully inject DI water into each section of tubing is a quick and easy way to look for the source of the problem.  Maybe a section of tubing is plugged, maybe it’s cracked, maybe there’s a dust bunny lodged in the end (true life this has happened to my IC!!)  It’s so hard to tell sometimes by just eyeballing the sections, but this technique has yet to fail me!

  • Look Beyond the Science

Sometimes the source of an instrument issue isn’t related to the “science-y” components we would typically focus on.  Sometimes exhaust fans wear out or autosampler screws fall out or sheer off.  Don’t be afraid to look into the more mechanical parts of your instruments.  Think of them as support systems.  If your autosampler syringe housing is loose and wobbly because a screw is loose, maybe the syringe isn’t being held firm enough to puncture the vials.  If you’re lucky, your instrument will be smarter than you.  Sometimes the software might notice that something is wrong and stop the analysis run.  If it’s not….you’ll come back to a broken syringe plus a lack of data.

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If you have any fun or interesting troubleshooting tricks that have never steered you wrong let me know!  I’m always on the hunt for new ideas, and if I get enough, we might do a part 2 later.  Having as many tools in your troubleshooting tool belt as possible is so beneficial, and it’s one of the areas that we should all constantly be striving to improve.

Part 3: How to Know if Your Calibration Curve is Correct

You’ve bought some standards and you made a multi-point curve.  You might think your work to improve laboratory accuracy is over, but then doubt starts to creep into your mind.  What if something’s gone wrong!?!  What if you run a sample and you expect one answer and your instrument tells you another?!  Is the sample bad, or is there a problem with your calibration curve?  Has something gone wrong somewhere in your plant’s process, or do you have a bad batch of calibration standards!?

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The first step in ensuring that your calibration curve is up to par is checking the Correlation Coefficient.  I know, I know….more math….but stick with me here because it’s important!  The R2 value of your calibration curve has a large amount of statistical power.  No one wants to talk about statistics, but it has a lot to do with how well your instrument is calibrated.

In a nutshell, the correlation coefficient gives you an idea of how well the standards relate to each other.  R2 values statistically can range from -1.0 to +1.0, but on your instruments, you should look for a value as close to 1.0 as possible!  Values of R2=0.9987, R2=0.99943, or R2=0.9963 are examples of what you are looking for.  The closer to 1.0 your R2 value is, the stronger the relationship between your standards.  A strong relationship between standards leads to more confidence in the reported results for your samples.  Remember that your standard value points hug your samples, and no one likes crappy, wet noodle armed hugs! It’s listed somewhere in your instrument’s software, you might just have to look around for it.

So you’ve got a strong curve, but calibration curves can be strong….but wrong!  How can you check the accuracy of the standards you used to make the curve?

There is a way to prove that you standards and instrument are operating correctly.  We are going to employ the Dr. Sheldon Cooper of standard….the validation standard!  Just like Dr. Copper, the validation standard knows more than you, and it isn’t afraid to tell you when you’re calibration is wrong!

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The validation standard should be a standard SEPARATE from your curve!  Buy it from an alternative supplier, buy a different lot of a standard you use in your curve, or buy 4 standards and just pick one to use only as a validation….just don’t use your validation standard as part of your calibration curve.

If you just analyze one of your calibration standards again as a sample, you will get the right answer 100% of the time….because somewhere in the software you’ve entered those values in as the answer.  This might look promising, but it tells you ZERO useful information about the accuracy and correctness of your calibration curve.  The validation standard’s “job” is essentially to be an unknown sample, that you secretly know the answer to.

A good basic rule of thumb is that the difference between the known value of your validation standard and the instrument’s reported result based on your curve should vary no more than 10%.  We’ll call this “Validation Recovery” for ease of terminology.  For most applications, 10% is much larger than the recovery should be, but if you’re just started down the path of improving your laboratory accuracy….it’s a good basic place to start.

From here we are going to have to dive into a bit more math, and I think we can all agree that one math topic per post is frankly too much math!

Next week I’ll cover the basic statistics of calculating a more precise validation recovery range for your instrument, tracking validation recovery, and how you can use that information to access the performance of your instrument!

Part 2: What Creates an Accurate Calibration Curve

Welcome back for the second week of Common Calibration Conundrums and Other Laboratory Queries!

 Before we get back into the science fun, I have an exciting announcement.  Bion Sciences now has an Instagram account!  Come follow along with us!  There will be blog notifications, access to our website, and photos of the Bion Sciences team in our natural, laboratory habitat!

@bionsciences605

 Part 2: What Creates an Accurate Calibration Curve

Now that we’ve covered how often you should calibrate an instrument, we should talk about how you pick standards to use for your curve and how many standards you should be using.

Think back to algebra when we first learned the equation for a line.  If your math teacher was anything like my math teacher, they drilled into your brain that 3 points are needed to accurately verify a line and the corresponding slope.  One point is simply a dot in space, and two points allow for too much swing in the correlation of the line, but three points……three points is really where the magic starts to happen!

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Don’t get overwhelmed with the math, I’ll simplify this for you!

The accuracy of your line, and therefore your results, improves dramatically as more data points are added to the line.  Think back to our puppy from last week.  If you only tell the puppy once a day, every day to potty outside, your results might be somewhat….questionable.  Now imagine you tell that puppy 3 times a day, every day, to potty outside.  The chances that puppy is going to catch on greatly improve!  The same goes for your instruments.

When it comes to calibration points, more is always better.  Again, there is no such thing as over calibrating an instrument!

The more data points you provide the instrument, the accuracy of the answers the instrument provides you will also improve.  The only issue that arises from more calibration standards is the time it takes to analyze each standard.  It’s important to find a balance between the most accurate curve you can produce and managing the time restrictions inside your lab.

To deal with time restrictions and busy schedules, sometimes improving your accuracy might involve decreasing the number of calibration points.  (Cue massive shock and awe as I appear to contradict everything I’ve said up to this point! 😊)

Let me give you an example.  Suppose you run 10 calibration standards….but because it takes several hours to build that impressive calibration curve you only calibrate your instrument every other month.  By the end of the second month, how true do you think those 10 data points are….I would argue they probably aren’t accurate anymore.  Instead, what if you ran 5 points once a week, or 3 points every day??  A smaller, more current curve will almost certainly produce more accurate data.  In the world of the laboratory, accuracy is the name of the game!

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So, you’re staring at a list of calibration standards and you find yourself wondering, “How do I figure out what to order?!”

A good rule of thumb is that your calibration standards should “hug” your expected value.  For example, if your anticipated answer is 3ppm, you wouldn’t want to make a curve using points 0.5ppm, 1.0 ppm, and 1.5 ppm.  You also wouldn’t want to use standards that all have values above your expected answer….meaning that 5.0ppm, 8.5ppm, and 12.25ppm also wouldn’t make a good curve for a value of 3ppm.

For an expected answer is 3ppm, I might select standards with values of 1.0 ppm, 2.5ppm, and 5.0ppm.  Typically, the wider the range of expected answers, the larger the range of standards should be.  Remember, your values need to be hugged by calibration points, not left to fend for themselves on the outskirts of your curve.  Every once in a while you might have a stray value that exceeds the boundaries of your calibration points, and that’s fine….every once in a while!  On the day-to-day and with your typical samples though, your results will be most correct if they are contained within the confines of your calibration data points!

Next week will be, Part 3: How to Know if Your Calibration Curve is Correct.  It’s Big Bang  Theory-themed so be sure to keep your eyes peeled for that!  In the meantime, if you have any questions about this week’s post or any ideas for future posts but sure to leave a comment and let me know!

Learning to Love Yourself

You wouldn’t know it by the weather this week, but summer time and bikini season is looming.  Usually, I’m always amped to throw on a swimmy suit and hit the beach or the pool, but this year, something is different…..

I think I’ve given myself unrealistic body goals.

I believe the root cause is in the fact that I sort of inadvertently lost so much weight last summer.  While struggling with and trying to get my anxiety and panic attacks under control last summer, I basically quit eating.  It wasn’t that I didn’t want to be eating, I’m not one of those girls who doesn’t eat…..food is always my jam!  I was just way too worked up all the time.  Eating just a couple bites of anything would just make me feel sick and nauseous.  I remember distinctly sitting at work trying to eat a cliff bar…..it took me 4 days to get through the whole bar!!  During this period I was also working out a ton, because ya know….that’s what they tell you to do when you’re stressed.  Run a bunch, lift all the heavy things and put them down, and get those happy endorphin feelings flowing!  I’m not sure how exactly I managed to function, but apparently anxiety gives you super powers….just not the sort of super powers anyone really wants or should have.

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It turns out that loosing 15 pounds in two weeks has a dramatic effect on your muffin top.  It also has a dramatic effect on your psyche…

I know that I only achieved my slimmed down self because I was unhappy, and that unhappiness and stress changed my body in ways that I normally cannot and would not do.  The worst part though….is that I felt so accomplished.  Maybe it was just the sense of control I had over things.  Nothing felt good, but I had finally achieved the look I had been craving since high school but never had.  It was the one little thing that seemed to be going in my favor.  I finally had reached a point where I didn’t have to squat stretch out my freshly washed jeans, I could throw on anything in my closet and there wasn’t a lump or bump to be found, and I think it just gave me this inflated and superficial sense of self-esteem…..which when you talk about it sounds super awful and unhealthy……BECAUSE IT WAS!

My body is obviously not meant to existing in a muffin top free state….I’m honestly not sure that even the strictest diet would have kept me looking that way.  Once I started feeling better the muffin top slowly but surely started making a come back.  I’m naturally curvy and just tend to carry and collect all my weight generally on my love handles and around my belly button…..a spot I’ve nicknamed “The Doughnut of Awful”.  I’m right around 5’10” so I’m overall very proportional and quite a healthy weight for my height.  I’ve never been told that I need to loose weight by any of my doctors, and even in high school at my most active, swimming and dancing for several hours everyday, I still was no where near as slim and lean as I was last summer.

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…..this photo stresses me out…..as much as I hate to say that and I want to rise above it all…..it still does.  I know that it’s only 12ish pounds and some summer time tan, but I’m not sure that this picture makes me feel as happy as the other one, which is my continual struggle.  There’s some muffin top and tummy squish, and my ribs and hips aren’t poking out of my body.  Idk….it’s not bad, I can rationalize that, but at the same time I don’t exactly feel good about it either.

Lately, I’ve been trying very hard to convince myself that a slightly squishy Miss Happenings is a happier Miss Happenings.

  • I try and remind myself that I’m in a happy relationship, and if I eat out a bit more or drink a bit more now than before it’s because I’m with someone who wants to spend time doing things and having fun with me.
  • I should be happy that my brain finally realizes that skipping a workout to have lunch with friends IS NOT AN ANNOYANCE OR AN INCONVENIENCE!!!  That’s just how my anxiety made me feel about it all the time.  It’s a rare chance to spend quality time with some of my favorite people.
  • I should realize that I rarely hang out in bikinis alone, so even if I’m looking slightly squishy in a bikini I’m still surrounded by people who love and care about me regardless of my muffin top.

I do honestly remind myself of all these things and more when I look in the mirror and don’t like what I see.  I suppose that I probably will always occasionally struggle with my weight and how I look.  I always have from time to time, so it doesn’t make sense that it would mystically go away anytime soon.  The most annoying and hardest part is the small little diva voice in the back of my mind that sometimes quietly whispers that if I just stopped eating for a couple weeks I would slim right down again.  Ugh….go away crazy diva voice…..I have burgers and shit to eat, new beers to try, and things to do that have nothing to do with you!

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Going forward, the best I can do is continue to work out, maybe reign in the diet and kick up the cardio a bit, and just generally live the best and healthiest life I can live!  You only get one life and one chance to do all the things you want to do.  I refuse to believe that anyone looks back on their life and regrets the fancy desserts they ate, or the summer time patio drinks with friends they drank.  That simply can’t be reality…..and if it is, I don’t want it to be my reality.  If I had continued on the stressed out path I was on, I know looking back that I would have regrets.  I already regret the things I did or didn’t do because my anxiety controlled my life.

I lived that life once, and I refuse to go back there.

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This summer, in the same jeans as last summer.  A little bit squishier than last summer, but I’m happy!  Ignore the sass face, The Engineer was mocking my photo skills….but he’s just jealous of my shooting skillz!

Happy is healthy….and that’s all there is to it!

 

Chapter 11: Math has Never Been My Friend

I realize it’s been quite some time since I have written about any of my dating escapades.  There hasn’t really been a particular reason for this, other than the fact that I guess I haven’t been going on as many dates!?!?  I’ve long since given up on all of my online dating apparati.  (Apparently the “correct” plural of apparatus is either ‘apparatuses’ or ‘two pieces of apparatus’……I personally think apparati sounds better so that’s what I’m gonna go with!)  I believe this is the first blog about a guy I actually had a long relationship with.  Of course, long is a relative term, but since all the relationships you’ve read about how been flings or short lived, I’ll consider this to be long.


 

Ahhh college, it’s a great time.  However, it’s also a time where you might make some questionable life choices.  I will be the first to admit that college me made some truly questionable choices.  Probably one of the most questionable……was ADD 3.  I refuse to look back on experiences with regret, because they have brought me to the life I live now.  Despite all my “middle class white girl struggles”, I really am loving my life right meow!!  Honestly though, if I knew that I would end up in exactly the same place I am now if I could just delete those two years from my life….I would.  I’m struggling with how to write this, because it could be quite long, but also because the majority of it was not good or healthy in any way.  I’m going to try and provide just enough background that you can grasp the situation I put myself in, and then I’ll try and just focus on the positive things I learned.

move-on-quote-relationship-text-true-Favim.com-274768Graduating college on the heels of an economic recession isn’t great.  You’ve put in tons of time, money, and energy, and done all the things you’d always been told you were supposed to do to be successful.  The ending result of all this time and money was that I was unable to find a job, despite all my best effort and hours of filling out seemingly identical applications.  I was firmly committed to the belief that moving home would be an admittance of defeat, so I did the next best thing I could think of….I moved in with ADD 3.  I knew the sort of jobs that were awaiting me back home…..essentially nothing.  I wasn’t willing to admit that sort of defeat, so against the wishes of my family….and probably my friends….I soldiered on with my plan.

After about a week, I didn’t want to be there!!  I had already known that ADD 3 had a rather substantial collection of tubs in his basement.  One of the perks of dating a guy with a house in college was that I got to do laundry for free….in the basement…..with the tubs.  Being the nice, unemployed, “housewife” that I was, I thought I would organize the basement!!  Most where garbage……random things….some of his old Army stuff…..and then I found them, TUBS FULL OF VERY NSFW THINGS!!!!!!!!

…………………………………AAAAAAAAAHHHHHHH……………….!!!!!!  

Somehow, I let him convince me that it would be ok to stay, that we would work it out, and everything would be fine.  I think part of me gave in to him because I felt like I had worked so hard and upset so many people by moving in with him in the first place that once again….I was unwilling to accept the defeat that would have been moving out.  Something about college me was apparently really into winning at this point….

I eventually got a job where I traveled frequently, so I was around less to put up with his crazy.  Yes….he was crazy.  Granted, I only know ADD 3 as a PTSD suffering Army veteran. I have no idea who he was or what he was like before he served our country and saw the things he saw.  In that sense, I have only the greatest amount of respect for him, his family, and the sacrifices they made.  He discovered that he could win any argument by playing the “Army Card”.  Honestly, there was never anything I could say or do other than sit there and take whatever random insult he had to throw my way because I could never bring myself to say anything back when he’d play the Army card.  You have to remember…..I was younger and stupider then……I’ve grown up a lot….he’s partly to thank.

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This basically continued for a year, until he decided to move to Florida….and I was so relieved.  I packed up all my stuff, and we drove it to my Dad’s house in Sturgis and honestly….I was happy!  I was happy he was moving away, and I was happy that I felt like I had finally found an out.  I really thought everything was going to change for me.  It did…..but at the same time, it didn’t.  I saw him twice more after that moving day in August…..once when he flew to Minneapolis for my birthday that December, and once in the Spring.  I….the girl who was so ready to let go of the relationship, just let him continue to have one with me.  I didn’t really encourage it, but I didn’t stop it either.  I suppose that’s partly to do with being young, and partly because those who are in bad relationships always seem to have a really hard time getting out of them.

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Ok….enough of this seeming like a pity party for me….that’s not at all what I want.  Again, I don’t regret it.  I survived, I’m stronger, and I’m the person I am today because he taught me I didn’t want to be the person he thought I was….AKA a spoiled rotten, Tinker Bell obsessed princess who would never survive on her own because I knew nothing about life.  He had a few positive impacts on my life, and that’s were we will end this….the good things.

  • My relationship with ADD 3 taught me that if you set aside your pride, your family and friends will always be there to support you…..no matter why you think you don’t deserve it
  • He bought me my glorious pupperoni Scout, who has been with me through everything for the past 5 years
  • He introduced me to my fabulous hair stylist who I still drive 6 hours to see
  • He taught me how to play poker…..kinda
  • When I’m told I can’t do something or I’m not worthy of something, it just pushes me to prove them wrong.  So……I guess I should thank him for my new car, paying my own bills, and everything else he ever told me I couldn’t do
  • He introduced me to Korean food…..which is delicious
  • Spicy hot ramen is now a favorite lunch food for one of my UBS girls because of him
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The night we brought my little Scout-a-rooni home from the shelter.  #Adoptdontshop