final thoughts!~

I can’t believe my time at Ecovative has come to an end!! It seemed like just yesterday since I first stepped foot and met my mentor. Through this STEM program, I have learned soooo very much both junior year and senior year about the science field and as I go into college, I feel much more prepared and ready to tackle my science labs than ever. This experience that I have received has helped me grow mentally and physically as an aspiring science major. There were times where I did not understand a single thing my mentor would tell me to do and I felt embarrassed and incompetent, but through all this, I was able to escape my fear of asking what I consider "stupid" (but not in reality) clarifying questions to my mentor, professor, teacher, etc. Both Courtney Hart and Christina were the best mentors I could have ever asked for as they have showed me my hidden capabilities in science! As I go to college, I hope that I will be able to find as great of internships as I was fortunate to have been offered this year and the last year at Emma. In addition, I am also grateful for the experience of being able to talk and share my work with my teachers (at the faculty meeting and at the end-of-the-year presentation). It was a more nerve-wrecking experience than presenting in front of my peers/under-classmates, but again, through it all, I have conquered one more fear and I actually found that I enjoy sharing my knowledge and experience with others.

All in all, I am extremely grateful for having been given the opportunity to explore my passion in the science field and helping me figure out what I want to do with my life after Emma academically! But most of all, I am very grateful for the STEM internship program as it has given me the confidence to pursue the science field despite the difficulties I have had throughout my high=school life academically with my science classes!

04/16/14

Last Wednesday, I was yet again very busy as there was a lot my mentor had planned for me. The highlight of today however was that she took me on a field trip to RPI. There we entered the lab as there was a particular very expensive machine (blanking out on the name) with which she wanted to run a filled test tube. This took me back to my junior internship with capillary electrophoresis. Put simply, she explained to me that within the test tube were certain protein strands and that this machine would churn out a graph filled with peaks indicating the particular proteins. This is my mentor's latest project and I believe she is aiming to label and find out the particular proteins that are beneficial to mycelium growth. The more she explained to me, the more I found out that this was not simply a matter of putting the test tube in the machine and getting automatically useful results. After the graph was printed, she showed me the various peaks and explained that not all of them are useful and that she had to separate the "noise" from the "peaks". Or in other words, only the very noticeable high peaks indicated protein strands while the frequent zig zag looking peaks at the bottom of the graph were useless information created by the default of the machine. In addition, I learned that even if she found seemingly a useful protein strand, she would have to analyze the peak and attempt to match the data of the peak (obtained from the data of the graph) form the data base of proteins and the peaks they make. This truly was a very tedious task and I was awed at how fast she came to matching the peaks of the protein with their names. She told me that after a while, one begins to become familiar with the different peaks and their particular characteristics. Furthermore, what brought false hope were the occasional really high peaks that I thought to be a high level of a particular protein, but found out to be levels of plastic from the test tube bottle--this my mentor told me was inevitable.

All in all, this was something different than my everyday working with touchable large substrates (as this was dealing with small portions of liquids and analyzing data--like that of my internship last year), and it was surprisingly nice to return to old experiences. I learned that rather than investing in large and extremely expensive machines like these, it was far better off to pay RPI money per hourly use until confirmed that this machine was truly beneficial to the further advancement of the company. (It was something like $150 an hour, but taking into account the actual price of the technology (which is probably several hundred thousand if not more...) this is the cheaper option.

Considering that I will be dealing with these types of machines all throughout college (as a biochemistry major), it was nice getting acquainted with the technology beforehand. :)

04/09/14

I got to work with Katie today! :)

On April 09, both Katie and my mentors were occupied and so we had the pleasure of working for Sarah--who also works in the lab at Ecovative. Ironically, today was the day that both Katie and I were the busiest as Sarah had a whole list of things she needed completed. Thus, this wednesday we were given full rein--or in other words, we were basically given a list of tasks and left to complete it by our selves. Had this been the first month of my internship at Ecovative, I would have completely freaked out, but having worked here for a lengthy amount of time, to my great surprise and delight, I knew precisely what to do and where to find the needed equipment--without my mentor anywhere near me. It was as if I was one of the working staff at Ecovative, moving from one facility to the other. (ie: the Lab to the Dirty Room and so forth).

I began my internship with a simple yet big task. I poured about 85 agar plates as their stock of these plates were running low. Although this was a rather simple task, I knew I was being of great help, or rather, I would like to think I was of big help, as no one here actually seemed to have the time to pour plates as they were off at meetings, running experiments, writing up reports, and the sort. These were simple PGA plates (aka peptone glucose agar plates). These plates are widely used for general cultivation of a wide variety of microorganisms. Glucose Peptone Agar is a highly nutritious medium that is able to support the growth of many fastidious microorganisms.

These are all the plates I finished pouring!

Me in action! 

After this was completed, Katie and I teamed up and

went to work in the dirty room. We put together about 

12 bags of a certain substrate (maybe even more!) and

prepared it for inoculation in the pressure cookers.

   

04/05/16

As I mentioned in the previous post, the experiment concerning floral foams with the clear flour came to an end as the company that Ecovative was testing for did not place a strict requirement/ priority. Consequently, today I started a new experiment but for the same project (dealing with floral foams).

The experiment deals with the concentration of surfactant. "A surfactant is briefly defined as a material that can greatly reduce the surface tension of water when used in very low concentrations." The surfactant used in particular for our floral foams is SugaNate. This material is very expensive and I was informed that it costs approximately $1.75 per block of floral foam using using only 0.1% concentration of SugaNate!!!. Thus, this test was to figure out the least amount of SugaNate required for floral foams still to perform their function. We tested concentrations of: 0.1%, 0.05% and 0.01% SugaNate. In short, what we are going to be looking for was how the uptake of blocks change with different concentrations of SugaNate.

The basic procedure is:
Suganate is incorporated into the Floral Foams via the pressure cookers. First, for approximately 1 hour, the block of floral foam is put in the cooker with NaOH in order to sterilize the foam (basic procedure). After this step, the block is moved to a different cooker with a mixture of SugaNate and water for about 5 minutes.

Hopefully, this coming Wednesday I find that 0.01% of SugaNate can still be used to produce similar results to that of 0.1% of SugaNate which will save Ecovative a lot of money!!

http://www.chemistry.co.nz/surfactants.htm

03/12/14

During my time at my internship, I was informed of the results that were collected from the ongoing project with the floral foams. Of the myriad of different experiments that we performed with these very foams, the one I wrote in particular about was the addition of clear flour. There were 3 floral foams involved: #1 was our control, #2 was the one to which we added 32grams of clear flour, and #3 was the one in which we added (approximately) 94 grams of clear flour. When I state that a number of clear flour was added, I mean, more specifically, to say that clear flour was added in the regrind (the material that makes up the floral foams).

In addition, for those who do not know or are mistaking clear flour with the everyday flour used in baking: "Clear flour is the portion of the flour remaining after the patent flour streams have been separated. Clear flour generally contain a higher percentage of protein than the other grades, but the quality of the protein is lower." Or in other words, clear flour is cheaper than the typical flour found in markets and it contains a higher percentage/about the same (or maybe just a little lower but the difference is negligible). Thus, Ecovative is killing two birds with one stone by applying clear flour over regular flour.

My mentor alerted me that for #1, mold appeared on day 5; for #2, mold appeared on day 5 and had better feature resolution; for #3, mold appeared on day 4. Thus, it was concluded that about 32grams was the amount of clear flour that would give Ecovative the best results as compared to no clear flour (#1) and too much clear flour (#3).

This stated, my mentor could have performed more tests to further figure out a more precise amount of clear flour that would produce the ideal floral foam. However, the company that this very product is for was content with the number of days as with 32grams of clear flour, the floral foams, as stated prior, can last for a minimum of 5 days without any mold, and on top of this, because it has good feature resolution, we have found that the mold typically begins underneath--a place where customers will not on their everyday-basis look at. In addition, most flowers do not have that long of a life-span and so it is most probable to state that the a majority of consumers will throw away the product before mold is actually seen.

http://www.thebakerynetwork.com/baking-science

02/26/14-03/05/14

On this day at Ecovative, I learned how to work the ECA machine. (Electro Chemical Activation technology). Using this machine, I prepared for and carried out chemical treatments for floral foams, a product that is currently in the workings at Ecovative. Put more simply, Electro Chemical Activation is used to make ECA solutions by "mixing readily available food grade salt with water thereafter passing the brine solution through patented reactors...once inside the reactor, the brine is activated by way on an electrical charge and two distinct solutions are produced."

Put in chemical terms:

H20 + NaCl <=> NaOH + HClO
(H+ + OH- + Na+ + Cl- <=> Na+ + OH- + H+ + ClO-)

This machine was used because normally mycelium produces Acly groups that are hydrophobic. However, because we were out to create floral foams (a spongy foam that soaks up water and acts both as a preservative to lengthen flower life and a support to hold them in place), we had to make the very groups hydrophilic. Thus, by treating it with ECA solutions, aka, by using a strong base to deacylate the group, we were, put in layman's terms, "chopping off" the hydrophobic part.

To the left is a diagram of an Acly group. R simply stands for any chemical chain. The hydrophobic part of acyl groups is the double bond between the carbon and the oxygen molecule. Consequently, by using the ECA machine, I deacylated the chain and "chopped" that specific part out to make the product hydrophobic.

The solution I am pouring out here is the solution made from the ECA machine that will be inserted and sealed with the raw floral foams (in bags) as shown in the bottom picture. This is so that the deacylate reaction takes place, making the floral foam products hydrophobic rather than hydrophilic. 

a more close up shot of the required amount of solution for each bag (which contained 4  raw floral foams products)

After inserting the solution into each of the bags, the next and final step was to seal the bags using the Vacmaster as shown above. 

02/19/14

 Last Wednesday, for the first time in quite some time, I did not work in the "dirty room" but rather spent my time working in the lab. The week before I had made the compost and sterilized each of the bags. This week, consequently, I took these same bags and proceeded with the next step, which is to take apart the fully-mixed substrate which had been "glued" together by the ingrown mycelium. A new thing that I had not previously known was that after completely separating the substrate into bits and pieces, a certain tailored amount of flour was added to the mix. The flour was said to help speed colonization once the substrate we took apart were inserted into packaging molds.

The material I was working with last Wednesday and the week before was twine. My mentor and I found this to be a very tricky material to work the week before as we have to laboriously take apart every single clump of twine before inserting it into the bag of substrate we were making. Sometimes we would need something like 35grams of twine and that was a pain. With other materials such as corn husk, all I would have to do is measure out 35grams of the material, but with twine, I had to carefully pick out and come up with 35 grams worth of pieces (or take apart the lumps) of twine that were not clumped together.
The difficulty with twine did not stop from there. This week, my mentor and I found that of the 7 bags that we had painstakingly put together, about 3 to 4 of them had formed mold. Thus, long story short, we were not able to use them and they had to go to the trash. (such a waste, my heart broke upon throwing of throwing the very bags that I had spent so much time working on). My mentor explained to me that the very bags most likely became contaminated due to the process in which they handle the material. After we had made the bags and put them in the heat chambers for sterilization, the next step was to take apart the substrate in the bags and give them a thorough mix via gloved hands. This is when she suspects the substrate became contaminated. (due to the bacteria in the air--despite it being done in the sterilized lab). She informed me that this had to be done given the particular characteristics of twine. Unless someone comes up with a different way, this was, unfortunately, the protocol when dealing with twine. This incident had me realize yet again how susceptible materials are to contamination and consequently how important it is to be sterile and take that extra precaution no matter how silly it may seem. It also made me realize that there are times when the whole batch prepared a week before becomes contaminated and when scientists have to start again from ground 0.