Let's hear it for van Leeuwenhoek!!

This week I am going to give thanks.  We all need the give thanks to the pioneers that paved the way for us, the budding scientist, biologists and innovators.
 
This week I'd like to give a special shout out to a late 17th century pioneer, Anton van Leeuwenhoek, the inventor and first user of the microscope, also know as the "father of the microscope".  His inspiration came from a set of glasses used by drapers to inspect the quality of the cloth in the factory.  van Leeuwenhoek taught himself the ins and outs of lenses and the ability to change the magnification by grinding and curving the lenses that he was using.  The best magnification of its time, 270x the diameter.    Below is an image of his invention, the first usable microscope.  Without his many failed attempts and his one victory, we would still be in the dark. 
 
Microscopes have come a long way since then and we have become very dependent on the functions preformed by it. I have been very fortunate this semester to use a great microscope for my experiments.  It is called a Zeiss Primo Star with an attached camera system called Axiocam. I am able to choose what type of magnification that I need ranging from 4x to 100x (oil immersion) which gives me the ability to see everything that is happening to my Microbes as my naked eye is nowhere as good.

van Leeuwenhoek was the first person to discover and view bacteria in a single drop of water, for this, I thank you.  My hay infusion experiment would never be able to reach its potential without his dedication to an idea. 
I hope that I or one of my fellow inters find or have a major breakthrough in their career that we will be able to read about in the history books.  Trial and error equals eventual success.  Good luck everyone, the race is on.

Microbe Metropolis

Happy Valentines Day!  I hope everyone's feeling the love day :o)

So lets talk Hay.

I've been going back and forth between experiments with relatively good results until I had a microbe explosion, on day 21 of my Hay Infusion lab. Here is an excerpt from my journal:

Day 17:
Not such a good day for microbes.  I  took samples from my pond water as usual. One from the top, middle and two from the bottom as I needed to be sure.  Nothing.  I inspected every inch of the microscope slide, from cover slip end to end but not a single microbe to be found.  Just some hay, a decent size air bubble and some yeast.  I took a second sample of pond water from the bottom of the jar (where I've seen them before) but still nothing.  I added some DI water and a fresh handful and called it an afternoon.

Oh yea...I upgraded to a big jar!

Day 21:
WOW - What a difference.  Looking at my first slide from the bottom of my set-up, a microbe Metropolis has emerged.  Take a look at my sample at 4x magnification.....they're everywhere!  I had to call Josh and Matt over to check it out.

It was really an exciting day, I was losing hope in finding anything cool in the waters of Papago but I was pleasantly surprised.  I can't wait to take my next sample to see what new colony is going to flourish.

Matt gave me a great book...an oldie but a goodie!  Its falling apart but it has an amazingly in depth color chart that makes the identification process a lot easier.  I was able to clearly see that I had an abundance of Diatomes/Desmids (the long thin worm or tube like ones) and plenty of Amoeba. It was amazing to see them in their natural state under magnification.  Some of those suckers can swim!

Chloro..who?? Part two.

So as I promised, here is the exciting conclusion to the experiment that I have been working on, so I’ll be the only one in the dark!!

I was instructed my Matt to be very careful with this next step as it involved and known noxious chemical that has been made famous by many movie villains…Chloroform, Reagent Plus >99.8%.  This stuff is nasty, you know the chemically soaked rag that the bad guy uses to knock out his victim, makes my experiment that much more interesting!!

  I of course turned the fume hood turned on high and stood with the glass partition separating my nose from the fumes, as I carefully measured out a mixture of Chloroform, Petroleum Ether and Isopropanol Alcohol and put them all into a developing chamber.  I then placed my TLC plates (two plates in one) into the developing chamber, set my timer for 45 minutes and once again, turned off the lights while the mobile phase takes place.  The purpose of this phase is for components of the mixture to move up the TLC places at different rates as it completes the development of the Chromatography or the visible bands of Chlorophyll A (bright green in color), Chlorophyll B (olive green in color) and Xanthropyle (yellow in color) that migrate across the silica gel that covers the TLC plates.  Once the mobile phase is over, the plates are removed from the development chamber and dried in order to scrape each of the bands off and examine them for the actual content or concentration of chlorophyll in each plant.

Calculating the concentration of chlorophyll is a pains taking process of finding the absorbency spectra of TLC purified pigment bands.  The colored silica gel is scraped off and mixed with alcohol in a centrifuge tube, placed on a vortex, and spun down once again.  The resulting liquid is placed in a cuvette and is examined in a Spectrophotometer which is a device to measure light intensity at different wavelengths.  It produces light with a light source, and after the light passes through a subject, the light is diffracted into a spectrum which is detected by a sensor and interpreted into results we can use. 

Next week I'll go into my results and give you an exciting update on my Mini-city microbes from the hay infusion lab.  Have a great week everyone!