Showing posts with label Labs. Show all posts
Showing posts with label Labs. Show all posts

Monday, February 11, 2013

What diatomists do to diatoms

February 11, 2013 0
My research is on the diatom communities of the Everglades. To study how the communities respond to environmental changes, I have to identify and count each of the diatom species I encounter under the microscope. To do that though, the diatoms have to be stripped clean of any organic material and other 'junk' in the sample. The diatoms go through a harsh bath of acid and heat, until all that is left of them are their empty but beautiful cell walls. This is possible because diatom cell walls are essentially glass.

Beakers of diatom samples mixed with hydrogen peroxide and nitric acid boiling away on a hot plate. Different amounts of organic matter (like peat or plants) and inorganic matter (like sand or clay) result in different colors and reactions.

In the Everglades, the limestone bedrock adds a lot of calcium carbonate into the soil and periphyton. Some components of periphyton (especially mucilage-producing filamentous algae) attract calcite crystals like a magnet. The addition of calcium carbonate, an inorganic substance, makes Everglades periphyton a bit more difficult to process all the junk away. That's because inorganic things don't really dissolve with the normal chemicals we use to get rid of organic stuff. But we don't want to try to dissolve too much of the inorganic material either, because we don't want to damage the diatom cells! Everglades samples go through an intense process including sulfuric acid, potassium permanganate, and oxalic acid. Check out this animation that shows how Everglades periphyton is transformed into a bubbly and acidic concoction, then into a white powdery layer of clean diatom cells at the bottom of the beaker:




Here are some before and after processing images of periphyton and diatoms under the microscope:





Thursday, November 8, 2012

Fear of Failure

November 08, 2012 0

Do you see the tiny green film at the bottom of the beaker in the picture above?  This extremely small clump of green stuff is a diatom sample weighing less than 20 milligrams that I need to carefully process and guard with my life for carbon isotope analysis. If I lose any of this sample or ruin a procedure, I will likely have to spend several weeks waiting for a new culture to grow and start over again.


Thoughts that have been running through my mind recently include:
  • What if I lose any of my sample while transferring it between beakers?
  • What if it spills?
  • What if a piece of hair lands in it? (note: things like hair and dust can ruin stable isotope samples)
  • Despite how many methods papers I read, I still feel like I have no idea what I'm doing. What if I dry the sample too long, or add too much acid or completely miss a secret step that isn't described in any journal articles?
  • WHY IS THIS FILTER FALLING APART? WHY? NOOOOO!!!!! 

Failure and data loss can happen at all stages of the scientific process. Maybe a storm destroys your field equipment, maybe someone steals a field instrument to sell it for scrap metal, maybe a boater decides to move that heavy metal box they found that happens to be collecting your data, maybe your computer crashes and you lose months of data analysis... and maybe, just maybe, the worst thing that could possibly happen occurs: you give a horrible thesis or dissertation defense.

Although science (and academia in general) demands perfection, failure does occur. It's likely the reason why journal articles published in 2010 can be about studies conducted in 1998- some sample was lost along the way and work needed to be redone. As scientists, we are expected to quickly move on from mistakes, learn from them, and start over again, whether the starting over sets us back days or years.


For now, I will hold on to these backup "diatom leftovers" that probably aren't usable but give me a false sense of security.

Grad Students, Scientists and Researchers: What is the worst lab mistake or scientific failure you ever made? What was the strangest way you have lost data or ruined samples? How did you get over these mishaps?

    Tuesday, September 25, 2012

    Everglades fashion: clean clothes are out of style

    September 25, 2012 0
    Whether we are working in the field or in the lab, the things we have to wear to do our work make us stand out from the undergrad crowd at our universities. Below, I have listed the latest styles for Fall 2012 Everglades research:
     

    Field
    Tannin-dyed Pants 

    Above is a picture of pants that I wear in the field. These pants are naturally tie-dyed with streaks of grey and orange from various organic compounds floating around in the water. The key rule in Everglades fashion is to never wear anything nice or new, so these khaki pants that I’ve had since 9th grade are perfect for the field.
    Ambiguous Event T-shirts


    Again, nice new clothing is never worn in the field, so research is the perfect opportunity to get use out of bizarre free T-shirts from volunteer events and raffle gift bags. If you are caught wearing Ambiguous Event T-shirts while transitioning between the field and your home, however, you will be inevitably be questioned by people who intensely read your T-shirt. “So how was volunteering at an Art Festival in Fort Pierce 8 years ago? What is Envirothon?”

    Hiking Boots


    I have two pairs of hiking boots. If I have to be in an enclosed space with other people, the newer hiking boots are worn. The older hiking boots, shown above, permanently live outside of my apartment due to mysterious swamp smells and are worn only for the most mucky activities.

    Field Accessories: Floppy hat, 1-inch thick layer of sunscreen
    , machete
     

    Lab
    Old Oversized Stained Lab Coat 

    Despite the fact that all stock images of scientists on the internet show people wearing crisp, clean lab coats, this is rarely the case. The purpose of a lab coat is to protect the rest of your body from spills and splashes of chemicals and dyes, so lab coats rarely look clean. Since labs don’t contain washing machines and no one ever takes responsibility to wash the lab coats, they are also rarely ever washed…

    Closed Toed Shoes 


    As a Floridian, I feel very awkward and uncomfortable wearing closed toed shoes (aka not sandals/flip flops).  Closed toed shoes, however, are usually required in labs.  Since I feel like I have giant feet when I wear regular running shoes with jeans, I invested a whole $7 in the shoes shown above, which are probably as close to sandals as possible.
     

    Old Outdated Jeans

    These ultra-low rise super flare circa 2003 jeans just won’t give out for lab work. They have micro holes either from lab chemicals or from my dog chewing on them, but until they rip in half, I will keep wearing them in lab.


    The Communal Lab Jacket
    All labs maintain a temperature of about 50 degrees, and all labs have a communal jacket that is shared among the employees. The person doing the most tedious task usually gets to wear the jacket (“Amber looks so cold weighing samples”). Like the lab coats, the communal jacket hasn’t been washed since 1998.

    Reseachers: Are there any lab or field fashions that I missed that you would like to mention?  How do you feel while wearing your lab/field gear in public?  Do you have any lab fashion advice (quick-drying clothes, least awkward closed toed shoes, how to make the communal lab jacket smell better)?
     

    Tuesday, August 14, 2012

    Grad Student Life Beyond Fieldwork: Part 4 (Our Labs)

    August 14, 2012 0

    Today’s post is the last installment of my series “Grad Student Life Beyond Fieldwork.” Take a look at Part 1 (Classes), Part 2 (Teaching) and Part 3 (Our Offices) to read about other facets of grad student life. Part 4, the post you are reading now, is about the labs where we produce and analyze data.


    Some measurements we use for our research, like water temperature and salinity, can be measured directly in the field. Other more complicated things like “cyanobacteria community structure” involve taking field samples back to a lab (or multiple labs) for extensive analysis. Below I have posted a “virtual tour” of the lab I work in, the Microbial Ecology Lab at FIU, to give readers an idea of what grad students and scientists do other than walk around wearing lab coats. 


    Above is the door to the lab. It basically says, “Welcome! You are about to encounter biological materials, carcinogens, flammable materials and high voltage equipment! You should probably turn around.” All of these dangers are necessary to understand what microscopic organisms live in water samples from the Everglades and Florida Bay. Although this sign implies that you might die upon opening the door, our lab is generally safe as long as you use common sense and have a little bit of lab safety training. 


    Next we enter the lab. Note that just like my office, it has no windows. All scientists are allergic to sunlight.

    One line of research conducted in this lab is determining cyanobacteria community structure. While one could look under a microscope at a water drop and take a guess at what kinds of cyanobacteria are in it, genetic techniques have been developed to give a much more accurate picture of what species are in a water sample. Understanding cyanobacteria community structure and factors that impact it is important for Florida Bay water samples because Florida Bay often experiences cyanobacterial blooms. Below is some of the equipment and tools we use to determine community structure.


    This is the PCR (polymerase chain reaction) thermocycler, which amplifies segments of cyanobacteria DNA. An extremely small drop of a water sample and several reagents are added to tiny vials then placed in this machine. The machine then repeatedly heats and cools the vials so copies of the desired DNA segment can be made.

    How small of a scale are we talking about? Above are the pipets that we use. Notice that they are set in microliters.

    Not all laboratory equipment is highly advanced and specialized. While the DNA is synthesized, agarose gels are prepared using the microwave shown above. Since ethidium bromide, a probable carcinogen, is used in the gels, this is not the best place to heat up your lunch. 

    After the gel is prepared and solidified, we pipet 5-10 microliters of the amplified DNA into tiny wells in the gel (I am probably really good at the board game “Operation” after doing this hundreds of times). An electric charge is then run across the gel in the gel electrophoresis machine shown above, which moves the DNA down the gel. The distance the DNA travels tells us how long the segment of DNA is. With a few more lab procedures and hours of analysis on Excel, we can determine what species of cyanobacteria were in the original sample.

    FCE Grad Students: How much time to you spend in your lab? What are some things you like or don’t like about your lab work?

    Other Readers: Do you have any questions or comments about our lab work?  Do you have any questions remaining about grad student life (classes, teaching responsibilities, etc.)?