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.

01/12/14: Different Types of Mushroom Substrate

          This past Wednesday, I spent my time again in the "dirty room" aka the room where Ecovative keeps all of their substrates. A substrate is basically any substance on which mycelium will grow on. There is an endless number of waste products mushrooms can decompose and when cultivating mushrooms, based on the particular substrate one can obtain different characteristics. Each time I come to Ecovative, I, time and time again, become amazed at the fact that mycologists can literally use almost anything we think of as "trash/waste" and turn it into a viable product that, in a nutshell, can help save the Earth!!

For instance, mycologists use most of the major debris that comes from farming or forestry.

       Of course one can not simply pick a batch of twigs from the forest floor and start cultivating. Mycelium grows best on substrates when they have been broken down into smaller pieces, 1/4 to 2 inches. 

Some of the different types of materials that can be processed by mycologists include:

• bambo
• brewery waste
• cacao shells
• cacti
• coconut/ coconut husk fiber
• coffee beans, grounds, hills, & debris
• corn, corncobs, cornstalks
• cotton & cotton waste
• fabrics
• garden waste, grass clippings, & yard debris
• hair
• hemp
• leaves
• manure
• nut casings & seed hulls
• oils (vegetable & petroleum)
• paper products (newspapers, cardboard, money, & books)
• soybean roughage
• straws (wheat, rye, rice, oats, barley, etc)
• sugarcane
• tea, tea waste, leaves, & trimmings
• textiles
• tobacco & tobacco stalks
• trees, shrubs, brush, & wooden construction waste

Stamets, Paul. Mycelium Running: How Mushrooms Can Help save the World. Berkeley, CA: Ten Speed, 2005. Print.

In spirit of Valentines Weekend, here's a cute image for you!

http://cheezburger.com/7485785344

02/05/14

I could not go to Ecovative this Wednesday because we had a snow day.

(Also, this is a very general summary of the steps that is required to create the packaging product)

01/29/14

Last Wednesday at Ecovative I helped make bags in the dirty room. What I did that day is the very first step scientists here perform in order to create their final mushroom packaging product. The dirty room is the room that contains all myriad of compost materials used to create their mushroom packaging product. For instance, there is a container that holds different waste products like saw dust and chopped up corn stalks and husks. It was a bit like cooking—like following a set recipe. After I filled the required number of bags needed to be completed for that day with compost, I weighed out a specific amount of calcium carbonate, CaCO₃ and added it to each bag. This substance, CaCO₃ has the effect of enhancing the myclelial linear growth rate. After this step, I added about a half a liter of water into each bag and thoroughly mixed all the components together. These plastic bags and their contents are then sterilized in an autoclave for about an hour and then allowed to cool. Then the mixture is injected with small pellets of mycelium and left to grow. I may have explained this process once before, but this was my first time that I actually got to start from the very beginning, with nothing but the rare ingredients. The person that I worked with, Christina, my mentor’s partner, showed me the different results/physical and chemical characteristics that one would get from using different compostable materials. When with my mentor, Courtney I mainly worked with analyzing the many different products, today I had fun stepping out of the zone and getting my hands dirty.

This video above may help you gain a better picture of what I meant in my explanation above. 

01/22/14

It’s known that a tip-high gradient of Ca2+ plays a regulatory role in mycelial tip growth in fungi and so I wondered whether potassium would have the same direct results. Unfortunately, as stated in my previous blog post, I found that this was not the case—either because something went wrong during the process of my experiment or whether there is a scientific explanation. As I had predicted after seeing my results, according to “The Effect of Cations on the Growth of Fungi” by E.B.G. Jones and D.H. Jennings, “Sodium stimulates dry weight production at low concentrations but inhibits is at high concentrations.” This statement fits very well with my results as the set of plates that was inserted with a low concentration of potassium bicarbonate exhibited some growth as compared to the plate that was given an excess of the very cation. Consequently, in my opinion, it seems that the reason the set of plates with the greater amount of potassium bicarbonate did not grow at all is because if I had inserted too much of the substance into my agar mix.

While researching for this very explanation, I came across several other published papers concerning potassium bicarbonate and mycelium. More-so-than serving as a growth stimulator like Ca2+, it seems to be that KHCO3 is a very effective natural fungicide. It has been found that increased concentrations of potassium bicarbonate (KHCO3) can act as an alternative to synthetic fungicides. Thus, it was concluded that potassium bicarbonate was an alternative chemical agent for controlling in particular, a natural antagonist: R. solani AG 4 HG-I and S. sclerotiorum.

If I were a researcher at Ecovative that had the goal of researching about potassium bicarbonate and mycelium growth, I would continue to experiment with these two substances, trying to find the ideal percentage of the cation. This is what many scientists like my mentor do, in hopes of bettering their mushroom product for their consumers. However, I do not believe I will be continuing on with this project as my mentor has notified me that mold had started to form on the different plates as a result of my long stay away from the lab.

01/15/14: A New Year @ Ecovative!

This week, I went into Evocative and as usual continued to experiment on my own experiment. For those who do not remember what this was about, the purpose of this experiment is to determine the effect of potassium bicarbonate on the growth rate of the mycelium in plated agar culture. During my last visit, I made 3 sets of plates: one set was made with a high concentration of Potassium Bicarbonate (17.6 g/L), one was made with a low concentration of Potassium Bicarbonate (1.76 g/L) , and a control set (made with your every-day Potato Glucose Agar [PGA]). Thus, all combined, I made a total of 24 plates.

Upon measuring the plates, I have found that the mycelium placed within a medium made with a high concentration did not grow at all. In addition, I was also shocked to find that the mycelium placed within the control agar plates, in general, grew the most. Those in low concentration did how, but only ever so slightly. An added observation that I noted was that there was no mold on any of the plates—thus, I knew my results were not the effect of contamination. This made me come to the conclusion that it is highly possible that the amount of high concentration I used (17.6 g/L) inhibited the mycelium from growing at all. As of now, I do not have a clear reason why this is so and consequently, this calls for more research on my part which I am also excited about! I really love unexpected results like these and rather than being disappointed at my results, this unanticipated outcome is one of the very reasons why I love science and it’s factor of the unknown J!!

Starting from the top down and in the picture in the left (and from left to right in the picture above) are 2 high concentration, low concentration, and control plates.

Although due to the condensation that formed inside the plates the growth is not that visible, the results, as mentioned in my post were very surprising!