January 15, Starting My Experiment!

Today, I worked with Courtney to begin the process for my first experiment!
To extremely summarize my DoE, I am looking to see if a dark treatment on the grow-it-yourself oyster mushroom kits Sarah is helping to develop will affect the wet weight of mushrooms.  Many commercial growers colonize oyster mushrooms with a 24 hour dark treatment.  We want to see if this will specifically work on this product.  Through data of the mushroom wet weight, we will be able to determine if a dark treatment is better than a light treatment.  Who doesn't want bigger mushrooms?
Courtney and Sarah already prepared substrate which they inoculated so I was able to dive into the regrind and packing parts.  This step creates the shape of the final product, a block, and allows for the mycelium to settle into this shape.  While I worked on packing parts, Courtney told me some more about her research.  One project she is currently working on is finding a way to quantify mycelium.  To do so, she is working closely with chemicals such as hormones, and their effect on the organisms.

These are bags of inoculation that Sarah and Courtney helped me make.  These bags were given the light treatment.  

I packed what was in the bags into parts like this.  This is from a bag that is undergoing the dark treatment.

This contraption (rack, black pod cover, and duct tape) is where the dark treatment takes place.  

After packing the parts, Courtney and I began discussing possibilities for an experiments that I myself would be interested in pursuing.  She gave me ideas that involved topics from micro-nutrients to agar or liquid inoculation.  While I am not working on this light treatment experiment, I will begin to research and find topics for my experiment.  Ecovative has several mushroom publications which I am allowed to borrow and my mentors are helping me to find science articles to broaden my understanding of mushrooms.

December 11, Fluorescent Mushrooms and DoE

Today was split working with Courtney and Sarah.  Courtney took me to the old lab because we were going to look at hyphae, the cells that make up fungi, and the microscopes were located in the old building.  The lab was more than half the size of the old one!

From my reading I learned there are three types of hyphae cells: generative, skeletal, and binding.  Courtney stained a variety of fungi samples which we used fluorescent microscopy to view.  Beneath are some pictures of what we saw.  

Fungi grown in a liquid agar culture; the tendrils are hyphae.  I believe this one is skeletal hyphae.

Close up of the previous picture.

 

Hyphae without fluorescent light.

Hyphae from mold.

Hyphae from mold, the round circles you see clustered at the edges are spores!

You can also see the mold spores closer up here.  The round, circular blobs that cover the surface are the spores.

This is an image of a clump cell, a means for transporting between hyphae.  This image was taken on an RPI microscope.

 These images don't compare to seeing the real thing.
After spending a good deal of time looking at these samples, I met with Sarah to discuss my first DoE, Design of Experiment.  My first experiment is going to be helping with Sarah's research with the coffee grinds previously mentioned.  Over the break, I will be writing up my DoE, and when I return we will begin work.  I am going to be looking at the mushroom's reaction to light during its growth.  It's exciting to be applying my lab report writing skills to a real-life situation.

December 4th, Busy Day of Mushrooms

On December 4th, I began the day by learning to make substrate with Sarah.  We mixed a variety of nutrients with the organic material which we then sterilized in pressure cookers.
Then I proceeded to mix and pack substrate and mycelium into plastic square containers for the fungi to grow in.  It was messy business, especially when I transferred a liquid agar culture.
Sarah then proceeded to show me examples of contamination.  Some were very obvious while others not so much.  We actually walked by two people, working with some contaminated mushroom and it was not pleasant to smell.  
Since we finished early, Sarah gave me more perspective of her work.  She told me about a project where she was trying to make a replacement for floral foam which contains many harmful substances.  Then she gave me a mushroom set part of an informal experiment, with the coffee grinds mentioned in earlier posts, for me to grow.  The pictures beneath are of the pack she gave to me.  Hopefully the mushrooms will successfully grow because then they can be harvested and eaten.
Next time, I get to begin working on my DoE (Design of Experiment).

November 27

No meeting due to confusion in ride schedule.

November 13, Grain Inoculation

This meeting was spent signing a few papers and working in the lab and I worked with Sarah.  I now have my own Ecovative email and crocs to use in the lab. :)

In the lab, we focused on transferring the mycelium on the agar culture I created the previous week onto grain cultures.  I learned that using grain cultures is one of the most efficient ways for spawning.  Each individual grain provides more surface area for mycelium to grow on as well as provide nutrition.  Therefore, you can mass produce mycelium spawn.

Sarah and I used liquid inoculation techniques to transfer the mycelium.  This is a method where you combine  fragmented mushroom mycelium (agar mycelium broken down with blender) with sterile water.  That way when you pour this mixture into the grain, mycelium fragments are more evenly distributed.  Like all things concerning mushroom growth, contamination prevention was important.  Once again, I worked on my lab skills and being aware of contamination.  When the transfer was finished, we put the spawn to incubation.

Afterwards, Sarah briefly showed me the project she is currently working on.  She is trying to create a product that is a raise-it-yourself mushroom kit that uses old coffee grounds.

November 6th, Training Begins!

On Tuesday November 6th, we began my training process, so I worked with Courtney in the laboratory.  I began the first few steps to cultivating mushrooms.  The first step is preparation and pouring of agar media into petri dishes.  Agar media is essentially the food for the mushrooms (mycelium) that we cultivate.  Courtney prepared two agar medias ahead of time because it needed to be put into a pressure cooker for sterilization and prevention of contamination.  Then the majority of the time I spent pouring the agar into petri dishes.
This turned out to be the most difficult thing I had done the whole day, but considering it was my first time working in a laboratory and that I had no previous sense of contamination prevention, this was understandable.  I couldn't put my hand or clothing above any opening and I learned to organize my tools in a fashion that would make pouring agar into petri dishes efficient without contaminating them.  Afterwards, we cut out small squares of mycelium from another petri dish to put into my agar medias; thus began the growth of my mushrooms!
I never realized how critical sterilization and contamination prevention could be, especially after reading The Mushroom Cultivator I was was assigned.  It is critical to prevent contamination to produce a pure culture of mushrooms.  Next time I will continue the training process and learning to cultivate mushrooms.

October 30th

No Internship due to Sandy :(