Leonardo Biral

<hr/>

 Hi Everyone!


 My name is Leo and I am a high school student from Marin County, CA. I love science, especially biology, and plan on majoring in a biology-related field at university and then going on to medical school. I am really happy to join such a great community of people that share my passion of science and am really looking forward to seeing all the interesting, creative ways people use their Foldscopes. I am also excited to put my Foldscope to good use for some of my projects and hope to share some of my work with you all.


 Currently, I am working on a microbiology research project exploring
 
 Pseudomonas aeruginosa, Escherichia Coli,
 
 and
 
 Staphylococcus aureus
 
 biofilm formations and am trying to isolate bacteriophages that could destroy them. However, I am mainly focusing my efforts on finding a way to destroy
 
 P. aeruginosa
 
 biofilms as doing so could be used to help cure the fatal lung infections that cannot be cured with antibiotics as the biofilm state that the bacteria are in renders them very resistant to traditional antibiotic therapy. These lung infections are the main cause of death of Cystic Fibrosis patients so I hope to be successful in my research.


 The last 2 photos below are of the petri dishes that I took out of the incubator this afternoon. They were treated with an agar nutrient gel and a
 
 P. aeruginosa
 
 (PSA) lawn was spread over the agar. Yesterday, I placed different combinations of PSA, E. coli, and Staph that grew for 24 hours two days ago in the quadrants over the PSA lawn. This afternoon, I checked these petri dishes and noticed that there were small zones of clearance at the edge of the triple culture quadrant in the 1st petri dish, while the other cultures were unable to stop the spread of the PSA lawn. The situation was similar in the 2nd petri dish.


 Although this is my 1st set of competition assays, I was able to get some pretty interesting results. I think that there might be some bacteriophage release from the E. coli and Staph in the triple culture as a defense against the PSA’s spreading. Since the E. coli and Staph were exposed to PSA when they were in the triple culture, they had adapted to effectively defend against it so they were able to do the same when exposed to the PSA lawn.


 I am going to retry this experiment and then try treating the bacteria with subtherapeutic doses of antibiotic or changing the bacteria ratios. Once I start getting consistent results, I am going to start testing for bacteriophages that target PSA and, if I find any, I will try to isolate them. I will also gather samples from these petri dishes and use my Foldscope to try and get a close look at the bacteria to just see what is happening in their world.


 Anyway, I am just so happy to be a part of this community and cannot wait to see what everyone does with their Foldscopes.




 Sincerely,


 Leo




 <img alt="FullSizeRender" class="alignnone size-medium wp-image-14139 gcloudimg" data-attachment-id="14139" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1458251620","copyright":"","focal_length":"4.15","iso":"200","shutter_speed":"0.033333333333333","title":"","orientation":"0"}' data-image-title="FullSizeRender" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_14133/FullSizeRender.jpg" width="300"/>
 <img alt="IMG_1302" class="alignnone size-medium wp-image-14137 gcloudimg" data-attachment-id="14137" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1458233215","copyright":"","focal_length":"4.15","iso":"80","shutter_speed":"0.033333333333333","title":"","orientation":"6"}' data-image-title="IMG_1302" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_14133/IMG_1302.jpg" width="300"/>
 <img alt="IMG_1301" class="alignnone size-medium wp-image-14136 gcloudimg" data-attachment-id="14136" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1458233202","copyright":"","focal_length":"4.15","iso":"64","shutter_speed":"0.033333333333333","title":"","orientation":"6"}' data-image-title="IMG_1301" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_14133/IMG_1301.jpg" width="300"/>

First Post! &#8211; Microbiology Independent Project

Hello everyone,


 It’s been awhile since my last post as I
 
 had really been struggling with the Foldscope setup. My main problem was aligning the light hole w
 
 
 ith the lens, but after hours of frustration and tutorial watching, I have been able to successfully view and take photos of my samples.
 


 I thought it would be best to keep it simple for my first Foldscope exploration and decided to observe a leaf sample from a Peony plant. I set up the light module on the Foldscope, paired my iPhone, and began exploring.


 The photos are not fantastic. They could be a lot more focused, but one can see more crisp images of the leaf sample in the exploration video here:



<div class="jetpack-video-wrapper">
 
 <iframe allowfullscreen="true" class="youtube-player" height="563" src="https://www.youtube.com/embed/lt9M3-0gV_o?version=3&amp;rel=1&amp;fs=1&amp;autohide=2&amp;showsearch=0&amp;showinfo=1&amp;iv_load_policy=1&amp;wmode=transparent" style="border:0;" type="text/html" width="1000">
 </iframe>
 
</div>

 It is really amazing how one can still see the individual cells and the minute details of the leaf even with my lack of experience and usage of the low power lens. This really says something about the observational power of this little microscope. I am really impressed.


 <img alt="IMG_1430" class="alignnone size-medium wp-image-16538 gcloudimg" data-attachment-id="16538" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1464385633","copyright":"","focal_length":"4.15","iso":"250","shutter_speed":"0.066666666666667","title":"","orientation":"6"}' data-image-title="IMG_1430" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16536/IMG_1430.jpg" width="300"/>
 <img alt="IMG_1429" class="alignnone size-medium wp-image-16539 gcloudimg" data-attachment-id="16539" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1464374749","copyright":"","focal_length":"4.15","iso":"1600","shutter_speed":"0.066666666666667","title":"","orientation":"6"}' data-image-title="IMG_1429" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16536/IMG_1429.jpg" width="300"/>
 <img alt="IMG_1425" class="alignnone size-medium wp-image-16540 gcloudimg" data-attachment-id="16540" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1464374727","copyright":"","focal_length":"4.15","iso":"1250","shutter_speed":"0.066666666666667","title":"","orientation":"6"}' data-image-title="IMG_1425" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16536/IMG_1425.jpg" width="300"/>
 <img alt="IMG_1433" class="alignnone size-medium wp-image-16537 gcloudimg" data-attachment-id="16537" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1464388552","copyright":"","focal_length":"4.15","iso":"400","shutter_speed":"0.066666666666667","title":"","orientation":"6"}' data-image-title="IMG_1433" data-recalc-dims="1" height="225" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16536/IMG_1433.jpg" width="300"/>


 Now that I have, to a certain extent, sorted out my initial issues with the Foldscope, I will regularly share my explorations and give updates on my microbiology individual research project.


 Sincerely,


 Leonardo Biral

Peony Leaf Exploration

Hi Everyone,


 In my first post, I talked a little bit about my microbiology independent research project that I have been working on for several months now. My project’s focus was destroying
 
 P. aeruginosa
 
 biofilm formations with isolated bacteriophages.


 My original method (the one talked about in my first post) was to grow different triple culture combinations of
 
 P. aeruginosa
 
 ,
 
 Staphylococcus aureus
 
 , and
 
 Escherichia coli
 
 overnight and then plate them on a
 
 P. aeruginosa
 
 lawn. When I did this, I observed zones of clearance around the areas where I plated the triple cultures on the lawn. However, when I took samples from those areas, grew them overnight in nutrient broth, centrifuged, filtered, and plated them on a
 
 P. aeruginosa
 
 lawn, no zones of clearance were formed as shown by the picture below. I only noticed plaques formed on top of the lawn. This is an interesting result in itself as it shows that whatever is occurring with triple cultures is giving the bacteria the ability to successfully establish a colony on the
 
 P. aeruginosa
 
 lawn whereas if
 
 E. coli
 
 or
 
 S. aureus
 
 were placed on a
 
 P. aeruginosa
 
 lawn by themselves, they would be overrun by the lawn.


 This was not enough. I wanted to create a plaqueless zone of clearance on the lawn, which is a clear indicator of a toxin or bacteriophage. I did some research and found an interesting procedure that described how to isolate bacteriophages that target
 
 P. aeruginosa
 
 from a solid sample. The solid samples I used were soil samples from three different locations in my school’s garden: a plant bed, the compost heap, and a dry soil heap. I took 1.5 grams of a soil type and treated it with SM Buffer. I took about 1.5 mL of the liquid soil/SM Buffer solution and centrifuged it for 10 minutes and filtered the supernatant  with a 0.45 micrometer filter. I then took about 1 mL of this solution and mixed it with 8.5 mL of nutrient broth and 0.5 mL of an overnight P.
 
 aeruginosa
 
 bacterial culture and grew it for 24 hours. After 24 hours, I divided each of these cultures in multiple Eppendorf tubes and centrifuged them. I filtered the supernatant and placed it on a
 
 P. aeruginosa
 
 lawn and incubated the plate for 24 hours. This process was done for each soil type.


 I did this experimental process twice and I got very interesting results both times. The 2nd time I did the experiment, I clearly noticed a plaqueless zone of inhibition in the lower left quadrant where I plated the refined sample from the dry soil, which is an indicator for a bacteriophage or toxin effective against
 
 P. aeruginosa
 
 as a competing bacteria would establish a colony in the middle of the zone of inhibition. The large zone of clearance in the bottom right corner is the result of a positive control, Ciprofloxacin. Check it out right here.


 <img alt="IMG_1412" class="alignnone wp-image-16747 gcloudimg" data-attachment-id="16747" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1464278064","copyright":"","focal_length":"4.15","iso":"40","shutter_speed":"0.033333333333333","title":"","orientation":"6"}' data-image-title="IMG_1412" data-recalc-dims="1" height="513" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1412.jpg" width="385"/>
 <img alt="IMG_1413" class="wp-image-16748 alignright gcloudimg" data-attachment-id="16748" data-comments-opened="1" data-image-meta='{"aperture":"2.2","credit":"","camera":"iPhone 6","caption":"","created_timestamp":"1464278068","copyright":"","focal_length":"4.15","iso":"40","shutter_speed":"0.033333333333333","title":"","orientation":"6"}' data-image-title="IMG_1413" data-recalc-dims="1" height="499" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1413.jpg" width="374"/>




 In my first post, someone gave me a great suggestion in the comments and said that I should try to observe biofilms with the Foldscope, so now that I have a better idea of how to use the Foldscope, I decided to give it a shot. I took a sample from the area near the zone of clearance formed on the 2nd plate, made a microscope slide with it, and began exploring! I also tried making observations with my school’s microscope and took photos from both explorations. The 1st photo was taken from the school microscope and the 2nd was taken from the Foldscope.




 <img alt="IMG_1446" class="alignnone wp-image-16742 gcloudimg" data-attachment-id="16742" data-comments-opened="1" data-image-meta='{"aperture":"0","credit":"","camera":"","caption":"","created_timestamp":"0","copyright":"","focal_length":"0","iso":"0","shutter_speed":"0","title":"","orientation":"0"}' data-image-title="IMG_1446" data-recalc-dims="1" height="559" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1446.png" width="315"/>
 <img alt="IMG_1449" class="wp-image-16743 alignnone gcloudimg" data-recalc-dims="1" height="559" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1449.png" width="315"/>

<div>
</div>

 It honestly boggles my mind how a microscope that costs roughly $1 to make provides the observer with a better quality image than a traditional microscope and is more accessible as taking photos and panning around images is significantly easier. On the traditional microscope, you get a clear overall view of the sample, but the Foldscope lets me see that and the smaller details. The small “grains” visible embedded in the purple  areas that represent the extracellular biofilm matrix are the
 
 P. aeruginosa
 
 bacterial cells.

<blockquote>
 
 More Foldscope Images
 
</blockquote>

 <img alt="IMG_1447 (1)" class="alignnone wp-image-16745 gcloudimg" data-attachment-id="16745" data-comments-opened="1" data-image-meta='{"aperture":"0","credit":"","camera":"","caption":"","created_timestamp":"0","copyright":"","focal_length":"0","iso":"0","shutter_speed":"0","title":"","orientation":"0"}' data-image-title="IMG_1447 (1)" data-recalc-dims="1" height="474" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1447-1.png" width="267"/>
 <img alt="IMG_1449" class="alignnone wp-image-16743 gcloudimg" data-attachment-id="16743" data-comments-opened="1" data-image-meta='{"aperture":"0","credit":"","camera":"","caption":"","created_timestamp":"0","copyright":"","focal_length":"0","iso":"0","shutter_speed":"0","title":"","orientation":"0"}' data-image-title="IMG_1449" data-recalc-dims="1" height="474" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1449.png" width="267"/>
 <img alt="IMG_1450" class="alignnone wp-image-16744 gcloudimg" data-attachment-id="16744" data-comments-opened="1" data-image-meta='{"aperture":"0","credit":"","camera":"","caption":"","created_timestamp":"0","copyright":"","focal_length":"0","iso":"0","shutter_speed":"0","title":"","orientation":"0"}' data-image-title="IMG_1450" data-recalc-dims="1" height="472" src="https://storage.googleapis.com/microcosmosdelta.appspot.com/images/transfer_16720/IMG_1450.png" width="266"/>




 I am really really impressed by how amazing, practical, and minimalistic the Foldscope is. Absolutely amazing!


 Thank you everyone,


 Leonardo Biral

Leonardo Biral

Earth

Posts

First Post! – Microbiology Independent Project

Peony Leaf Exploration

Pseudomonas aeruginosa Biofilm Foldscope Exploration

Locations

Statistics

Type of Sample

Foldscope Lens Magnification