Tuesday, May 8, 2018

If you don't eat your bread, it WILL!





Each semester, the microbiology students make a visit to my lab to see the scanning electron microscope to get an idea how electron microscopy is done, or at least as much as they soak up in about 20 minutes.  During this time they are studying fungi so I use one of the (non-pathogenic) black bread molds as the sample for the SEM, in this case Rhizopus stolonifer. This semester I decided to make a try some new settings on the SEM and was able to make better images than usual so I thought I would show you what I have. 

[On a technical note my lab does not have a sputter coater, so I literally use a pair of forceps to transfer a small sample to the copper tape on the stub and put the sample right into the microscope, what I call down and dirty electron microscopy. The SEM in the lab is an Hitachi S-3400N.  I used the backscatter detector, working distance 4-5 mm, 70-80 Pascals, 10 kV, probe current 35-50, with the aperture set at 4]

Let's start with the sample.  These are cell phone camera images.


Fungi samples for microbiology are grown in Petri dishes containing potato-dextrose agar.  This shows the plate open, lid on the right. The culture was a couple of weeks old and seriously overgrown, but still great for my purposes.

A closer look at Rhizopus in the Petri dish.  You can see why it is called a black bread mold even though the hyphae - the fibers that make up the "body" of the fungus - are white.  The little black spheres here are sporangia.  Cumulatively, the sporangia make countless numbers of spores which spread the fungus to new habitats.


The next two images were taken with a Leica dissecting microscope.


This image is a composite of 28 images that were focus-stacked in Photoshop.  It shows the tread-like hyphae and the darker sporangiophores with sporangia at the top.  You can just make out the spores that make up the sporangia.

Higher magnification of the the same area as shown above.  Magnification here is 125x. This is a composite of 17 focus-stacked images.


So what the heck is focus stacking?  

The depth of focus for the lenses on a light microscopes is very shallow which means that if there is any z-axis depth to your sample only one tiny part will be in focus.  The image below shows one of the images used to make the previous image and illustrates what I am talking about.

One of the images used to make the focus-stacked image above.  Everything either above or below the focal plane is out of focus.  Even the large sporangium at left center is only partially in focus.

To focus stack you take a series of images, slowing moving the stage down (or up) after each image.  Ideally, the focus depths of sequential images should overlap.  Once the images are taken, Photoshop does the rest by selecting the part of each image that is in focus and combining those parts into a single image. 

As Arthur C. Clarke wrote, "Any sufficiently advanced technology is indistinguishable from magic."  I know this is actually very clever programming, but it sure looks "magical" when Photoshop reveals the final image.

Scanning Electron Microscope Images


One of the nice things about electron microscopy is that you don't have to focus stack.  The depth of field is much deeper and can be controlled.

In the image above the white, thread-like filaments that make up the fungus are obvious in the upper, right-hand portion of the image.  A mass of hyphae is called a mycelium. [SEM image; 55x] 

This is the sporangium in the center of the image above.  The stalk that supports it is the sporangiophore. [SEM image; 190x]



Note that the texture of the spores has become apparent as well as some small crystals of potassium chloride that have formed on the surface of some of the spores. [SEM image; 1,000x]

At 5,000x lots of detail shows up in the SEM.  



Note that the spore in this image has several holes in it.  Those were accidentally burned into the spore when I really zoomed in on it.  This is  called beam damage (dang it).  Also notice the details of structure the can be seen in this 15,000x image.
What causes beam damage and why isn't it across the entire sample?

One trick in electron microscopy is to zoom way, way in - even higher than you plan to capture - and focus.  As you back out on the magnification, the entire image remains in focus.  The SEM puts out a set amount of energy, regardless of magnification.  As you zoom in, the energy is concentrated into a smaller, and smaller area that can, when imaging something like a spore, begin to burn the specimen.

These holes are actually very tiny.  This is a 50,000x magnification image.  The SEM is calibrated so I can make measurements directly on the image.  This hole is 146 nm across or 0.000146 mm. Small.
This image, at 35,000x, shows one of the salt crystals on the surface of the spore.  It is 549 nm across. 

This last image, at 55,100x, is of a salt crystal that was on the side of a spore. I just thought it looked interesting, sort of like the head of a turtle.

One of the things I try to impress upon our microbiology students is that we humans are in a continuing competition with fungi for our food.  You probably don't think about it, but when you throw away old, out-of-date food or food scraps, ultimately, you are feeding the fungi.  It is very nice of you go to work every day to make the money to buy them food.  I am sure they appreciate it.  

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

All images in this blog are covered by Creative Commons license.  You may download, use, or modify the images any way you like just as long as you attribute the Eastfield College Microscopy Lab and don't sell them.

Best,

Murry Gans






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    +919164179356


    +919164179356

    +919164179356


    +919164179356


    +919164179356

    ReplyDelete
  7. IS IT POSSIBLE TO GET A SUCCESSFUL RECOVERY OF YOUR FUNDS LOST TO FAKE/ILLEGAL CRYPTO WEBSITES OR ANY ONLINE FRAUDSTAR/ FRAUDSTARS ? ABSOLUTELY YES!

    If you have had an encounter with any of the following fraudulent schemes;
    fake brokers, fake bitcoin investment platforms, forex trade, binary option, false investments proposals, you are reading the right post 👍🏻

    BLOOMBERG inc. Is a group of international cyber security specialists bound by one sole aim, which is to render state of the art services involving ethical hacking.
    Ladies and gentlemen I must warn you that the internet isn't quite a safe haven, as consistent research has found out that 62.24% of people you interact with on the net do not exist.Hence, you must trade with caution ⚠️ .
    For this reason, Bloomberg Incorporation was founded in the year 2005 and ever since have been trending for successfully carrying out various hacking jobs.

    Other services we offer includes;
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    Your happiness is our priority.
    ☠️Bloomberghackers@gmail com
    📌exploithackersinn@gmail.com
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    ReplyDelete
  8. ARE YOU LOOKING FOR A PROFESSIONAL GENUINE HACKER ONLINE❓HAVE YOU LOST YOUR MONEY TO SCAMMERS💵? OR DO YOU WANT TO CATCH YOUR CHEATING SPOUSE🚻? AND LOTS MORE ! THEN TRY DCLEAKS INC. WHERE EVERY REQUEST ON HACKINGS ARE SOLVED.

    DCLeaks was created in June, 2016. It consists of a group of GREY HAT HACKERS and is an affiliate to Russian based hacking organisations such as Evil Corp and Fancy Bears. We have our interest focused on fighting online scam and exposing cheating/corrupt individuals through the efforts of our qualified Hackers who are recruited and registered. We do this by making sure that our clients get the best services on a basis of PAYMENT AFTER JOB SERVICE✔️. You can test run and make sure that the job is done before payment is made.
    Our Customer service is 💯 guaranteed as we have received over 10,000 TESTIMONIES from all over the world and we offer one of the best Hacking services in the world.🌎

    Other services that we offer are listed below:
    ◾️Social media account
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    ◾️Android and iPhone hack.📱✔️
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    DCLeaks Inc.
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    Terms and Conditions apply.

    ReplyDelete
  9. Yes it is possible to retrieve your stolen bitcoins. I never believed in bitcoin recovery because I was made to understand that it is not possible. But sometime in February I fell for a binary options scam which promised a higher return and I lost close to $55,000. I read an article on here (reddit) as regards to a recovery expert and genius so I reached out to ghostchampionwizard, and to my surprise I got all bitcoins recovered within 42hours frame. I don’t know if I’m allowed to share the links on here but you can contact him if you are finding it very difficult to withdraw your funds from all this crypto investment scheme to recover your funds and your lost Btc. CONTACT HIM AT : GHOSTCHAMPIONWIZARD@GMAIL.COM

    ReplyDelete
  10. ARE YOU LOOKING FOR A PROFESSIONAL HACKER ONLINE❓HAVE YOU LOST YOUR MONEY TO SCAMMERS💵? OR DO YOU WANT TO CATCH YOUR CHEATING SPOUSE🚻? AND LOTS MORE ! THEN TRY DCLEAKS INC. WHERE EVERY REQUEST ON HACKINGS ARE SOLVED.

    DCLeaks was created in June, 2016. It consists of a group of GREY HAT HACKERS and is an affiliate to Russian based hacking organisations such as Evil Corp and Fancy Bears. We have our interest focused on fighting online scam and exposing cheating/corrupt individuals through the efforts of our qualified Hackers who are recruited and registered. We do this by making sure that our clients get the best services on a basis of PAYMENT AFTER JOB SERVICE✔️. You can test run and make sure that the job is done before payment is made.
    Our Customer service is 💯 guaranteed as we have received over 10,000 TESTIMONIES from all over the world and we offer one of the best Hacking services in the world.🌎

    Other services that we offer are listed below:
    ◾️Social media account
    hack (Facebook, Instagram, Twitter, Snap chat etc).✔️
    ◾️Recovery of Lost Funds to Scammers (Binaryoptions scam, bitcoin scam and romance scam).💵💰✔️
    ◾️Android and iPhone hack.📱✔️
    ◾️Cryptocurrency wallet repairs✔️
    ◾️Data base hack.💻✔️
    ◾️Credit card debt clearing.💳✔️
    ◾️Credit score upgrade.✔️
    ◾️E-mail hack.📬✔️
    ◾️Website hack.📡✔️
    ◾️Other Computer hacking related Jobs.
    Your Satisfaction is our Pride!
    For more information about DCLeaks, check DCLeaks (Wikipedia).

    CONTACT:
    ✉️ Worldeaglehackers@gmail.com
    Direct messages.
    +14433474376

    DCLeaks Inc.
    Copyright©️2021. All rights reserved®️
    Terms and Conditions apply.

    ReplyDelete
  11. I have been in a serious Situation where my Husband always comes home late at night and is always on his phone chatting on Instagram and Snapchat which I don't have access to his phone but I can't confront him about it either so I saw a Testimonial About Wizard Schwartz Hacker So I contacted him via:schwartzsoftwarehackingprogram@gmail.com who reveal many things about my husband he was able to hack his device and get a lot of proof for me but I have to go extra miles for him to hack his phone conversation so I can always listen to it as well, right now I can read his chat from my mobile device through a Hacking link he send for me if you are in such situation like me contact him now: schwartzsoftwarehackingprogram@gmail.com / +1 704-313-9661

    ReplyDelete
  12. Have you lost hope in trying to recover stolen funds from a scammer?
    Do you keep meeting scammers in search for an hacker to recover those stolen funds ?
    Are you curious about if he is cheating?
    Are you worried about what the kids do with their gadgets?
    Do you want to know what she is doing and where she is at all times?
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    ReplyDelete
  13. I want to express how grateful I am to CYBERCRDITGURU a credible and reliable hacker who helped recovered the $45,000 I lost to scammers online and also hacked into my cheating husband’s mobile phone and social media accounts. I am married with 3 kids although my husband is extravagant and cheats on me with a lot of side chicks, he hardly comes home. Then I saw this investment platform online and decided to invest the sum of $45,000 to it, with the intention to grow it but I never got my money back neither capital nor profit. “I was scammed”. However, I discovered CYBERCREDITGURU through positive reviews read on MerchantCircle and reached out to them immediately on CYBERCREDITGURU (AT) GMAIL DOT COM and phone/text: +1 (650) 439 0624 and they helped recovered my lost $45,000 and helped me monitor my husband’s calls and social media accounts. I now have my husband back. I can’t thank them enough but give them a 5-star review as a token of my sincere appreciation. I strongly recommend them to anyone with related issues. Here is there contact again: CYBERCREDITGURU (AT) GMAIL DOT COM and phone/text: +1 (650) 439 0624

    ReplyDelete
  14. CRYPTO INVESTMENT SCAM RECOVERY

    By now you should know about the Crypto investment scam. They are all over the internet posting Enticing adverts and making false testimonies about themselves using this to Lure people to their advantage and Scamming them, But you need not to worry, cause you can get all your money back.

    You might have also come across some individuals that say they will give you guarantee on successful trades but they only end up as SCAMMERS as well. You here them say stuffs like 200% guaranteed in just 2 weeks and when you go into trade with them, they start telling you to pay profits percentage before you can get your income. These are all liars please avoid them. But if you have been a victim of this guys, then you should contact PYTHONAX now!

    The big Question is “Can someone Recover their money lost to Binary Option and Scam!?
    I will say yes, and will tell you how.

    The only way to Recovery your money back is by hiring HACKERS to help break into the Scam platform Database Security System using the information you provide them with, Extract your file and get back your money. It seems like a really impossible thing to do, I will tell you, it should be impossible, but with the use of specially designed softwares known to HACKERS and Authorities (such as The FBI, CIA e.t.c) it is possible and the only way to recover your money.

    PYTHONAX are a group of hackers who use their hacking skill to hunt down SCAMMERS and help individuals recovery their money from Internet SCAMMERS.
    We just need the contact details of the SCAMMERS and Paymnet Info and within 4-8 hours your money will be return to you.

    You can contact us via the emails below;
    Help@pythonax.tech
    Hackservices@pythonax.tech


    Pythonax.
    2023 © All Right Reserved.

    ReplyDelete