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» LymeNet Flash » Questions and Discussion » Medical Questions » The Microscopy Thread (Page 13)

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Author Topic: The Microscopy Thread
bluelyme
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With amscope usb cam the resolution was good but the framerate was really slow ..i sped up frame rate and lost resolution. Gunna return it and try ccd to tv ...my friend had better results seeing ketes

unless you can get a trinocular dslr hd setup ...i want to monitor as i experiment .. tnt has a good rig with american optical..try phase or darkfield if you can afford it

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Blue

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BorreJaakko
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Thanks for the reply!

As I am from Europe, I would prefer something that is available here.

Now that I have read about the thing a little bit more, I think DSLR and some used microscope going with it is probably the way I want to go, as I was thinking about getting a DSLR anyway.

In my country there is only one suitable used microscope for sale it seems, somebody is selling an used Olympus BX-41 with only binoculars and basic optics, that is it. Asking price is 2000 euros. I don't know whether it is a ripoff or not. The price of Olympus BX-41's varies widely.

In any case I would need to get trinoculars or remove the binoculars and get some kind of adapter to connect it directly to a DSLR.

These guys seem to sell some ready made and custom adapters.

http://www.lmscope.com/produkt22/produkte_en.shtml

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bluelyme
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Looks highly adaptable. ..does it come with phase contrast or dark field ?

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Blue

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thatdudefromkansas
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I use my DSLR for all of my images and video.

However, my 70d doesn't do time-lapse without downloading new software, which I might do in the future.

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M L
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Hi BorreJaakko,

You can see Lyme spirochetes with a bright field microscope, but dark field ones (more expensive) are better for looking at blood. Plus, a dark field capable one is usually also bright field capable, so you can do both with the one microscope for different viewing options.

I have this one: http://www.amscope.com/40x-2000x-trinocular-compound-darkfield-microscope-with-oil-condenser.html?gclid=COWNwYjfsM4CFcdehgodgMgJKA#product_tabs_product_videos

I also bought a USB camera from AmScope (3MP USB2.0 Microscope Digital Camera). The 3MP one isn't as great as I would've liked (what I see with my eyes looking in the microscope is a lot more clear than the camera pictures/videos, but you can still capture and discern the specimens). Definitely don't get less than 3MP if you end up buying one.

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thatdudefromkansas
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ML, I use the same microscope, but upgraded the 100x objective to one with an adjustable iris.
https://www.youtube.com/watch?v=dH3fGVndMYo

Sample video using my DSLR.

Need to start getting to work on microscopy again.

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M L
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That's a great idea about swapping out the 100x for one with the adjustable iris, thanks. I couldn't get anything to focus with the 100x it came with, so I just used the other objectives.

That's a really great video!

I never even thought that a camera could have an adapter for a microscope - mind blown. Lol

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thatdudefromkansas
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Yep, I bought the adapter off amazon.

It works pretty well.

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TNT
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Hi fellows, it's been a little while. I hope all of you are doing well, or at least are ok.

I did a stained smear recently and am seeing things that really look to me to be Anaplasma. So, I thought I would put these up for you guys to see as well. I also have some cool pics of biofilm.

Clinically speaking, my symptoms and response to treatment very much suggest a Rickettsia. Also, BLO & Mycoplasma.

First, my biofilm pics:

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BLO-like organisms:

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The possible Anaplasma pics:

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The same with a different focus:

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[ 10-25-2016, 03:54 PM: Message edited by: TNT ]

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TNT
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A different neutrophil:

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Possible extra-erythrocytic Anaplasmal elemental bodies:

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Possible elemental bodies (small morulae) in a Neutrophil:

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Very possible elemental bodies in a platelet, or otherwise referred to as Anaplasma platys:

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Very possible elemental bodies in a red blood cell:

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I've been making steady incremental progress, and Levaquin is definitely kicking some pathogens' butts. Tetracycline, BVT, and Rifing are helping too. I'm not where I need to be (that's an understatement), but am making some progress.

[ 10-25-2016, 04:07 PM: Message edited by: TNT ]

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TNT
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I couldn't get tinypic to work right for me today, so I posted from Google. Let me know if you can see these pics.
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M L
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The pictures aren't loading for me on multiple browsers
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TNT
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That's what I was afraid of. Thanks. I'll try to correct this as soon as I can.

Sorry about that, guys.

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TNT
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Bringing this thread back up. The pics are now visible.
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M L
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Those are great pics- awesome job with the staining! Thanks for sharing and adding the specific captions. I hope you progressively keep getting better
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Lymedin2010
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That looks like anaplasma for sure, as they typically appear on the peripheral of the nucleus & close to the cell wall most times.

Great job!

I will share your photo with others too. Another person with Lyme also has identified Anaplasma in his blood about a year ago or so.

I am not too certain about the babesia, as they may still be artifacts from the many larger ones. Hard to tell for sure for me.

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jsnyde2
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My Hold the Mayo infectious disease doc did microscopy and found nothing. Why?
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birthdaysuit
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I'm looking into purchasing a microscope, my budget is 1000 dollars and under. I would like something that is clear and has an a adapter for a digital handheld camera. Not sure if a 60x optical zoom with 500x regular zoom handheld camera would work in this application or if something like a Canon DSLR 60d is better.

Basically, I want something that isn't useless and is able to show clearly the spirochete in serum. I'm leaning towards Dark-field but I understand past 400x is markedly expensive, well over 1000 dollars. I'm wondering if a camera would compensate? Lightfield, phase contrast or DIC are fine too but I'm looking primarily for a microscope, whether it be 30 years old or not that can see the spirochete clearly. I understand there are a lot of crappy microscopes out there not worth the money.

Thanks, I'm hoping for some input on this matter!

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TNT
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Thanks fellows!

quote:
Originally posted by Lymedin2010:
Another person with Lyme also has identified Anaplasma in his blood about a year ago or so.

Are there pictures you could post here? I would be interested in seeing them.


quote:
Originally posted by Lymedin2010:
I am not too certain about the babesia, as they may still be artifacts from the many larger ones. Hard to tell for sure for me.

I'm not sure what you are referring to, as none of the pics I just posted have babesia in them.
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TNT
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quote:
Originally posted by jsnyde2:
My Hold the Mayo infectious disease doc did microscopy and found nothing. Why?

Standard procedure for viewing stained slides is usually only a few minutes. It sometimes takes hours of viewing to see these things. Even Dr. S in Florida (who has multiple books about the co-infections) says this.

There's a fairly steep learning curve to doing this yourself (at least for me it was), but the only way you will get someone to look at your blood for longer than 15 minutes is to do it yourself.

Besides, Mayo has discredited themselves time and time again. The swath of carnage they have caused in relation to Lyme disease is horrendous.

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TNT
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quote:
Originally posted by birthdaysuit:
I'm looking into purchasing a microscope, my budget is 1000 dollars and under. I would like something that is clear and has an a adapter for a digital handheld camera. Not sure if a 60x optical zoom with 500x regular zoom handheld camera would work in this application or if something like a Canon DSLR 60d is better.

Basically, I want something that isn't useless and is able to show clearly the spirochete in serum. I'm leaning towards Dark-field but I understand past 400x is markedly expensive, well over 1000 dollars. I'm wondering if a camera would compensate? Lightfield, phase contrast or DIC are fine too but I'm looking primarily for a microscope, whether it be 30 years old or not that can see the spirochete clearly. I understand there are a lot of crappy microscopes out there not worth the money.

Thanks, I'm hoping for some input on this matter!

Welcome to Lymenet, birthdaysuit!

With that budget you should be able to get a pretty decent microscope. For doing digital captures it's very handy to have a trinocular. Adapters are microscope and camera specific, unless you get a universal one, like the one Lymedin2010 linked us to a while back.

Zeiss, Nikon, Olympus, American Optical, Reichert are good lab and research-grade scopes we recommend. Some here are doing excellent work with Amscope (like "dudefromkansas"), but he purchased a good 100x objective for his scope. The 100x objective his Amscope came with was useless.

You can see spirochetes in live blood with only 400x, but full 1000x is very handy.

We recommend a scope also equipped with darkfield, or phase contrast, or both. They make it very easy to see live objects without stain, but you can see live ketes with just brightfield, it's just more difficult.

To view Giemsa-stained slides all you have to have is brightfield, but you do need 1000x for that.

I bought my AO trinocular phase contrast scope for just over $100, but that was a really great deal.

Read over this whole thread, and you will get a better idea of what is involved with microscopy.

Here's another very informative forum I like:
http://www.microbehunter.com/microscopy-forum/

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Lymetowne30
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Hi
Newbie here. I really have no idea how I'm to post a question on here RE; my blood. I'm in desperate need of some options as my Lyme specialist doesn't know what's going on either. I have a video plus photos. Any help= much appreciated

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bluelyme
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Welcome ltown30,you can use tinypic or youtoobs for vid...

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Blue

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bluelyme
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I finally got a decent cam for my pos ...is this a filarial,it is more visable when rbc pass over it ?.
Tnt i will be doing some more staining to check for those infected neutrophils
i caught another one of weird bubbly sop? I will try to upload soon ....i guess more iver and art ...?

https://www.youtube.com/watch?v=5MLuVU_YGjo&feature=youtu.be

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birthdaysuit
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quote:
Originally posted by TNT:
quote:
Originally posted by birthdaysuit:
I'm looking into purchasing a microscope, my budget is 1000 dollars and under. I would like something that is clear and has an a adapter for a digital handheld camera. Not sure if a 60x optical zoom with 500x regular zoom handheld camera would work in this application or if something like a Canon DSLR 60d is better.

Basically, I want something that isn't useless and is able to show clearly the spirochete in serum. I'm leaning towards Dark-field but I understand past 400x is markedly expensive, well over 1000 dollars. I'm wondering if a camera would compensate? Lightfield, phase contrast or DIC are fine too but I'm looking primarily for a microscope, whether it be 30 years old or not that can see the spirochete clearly. I understand there are a lot of crappy microscopes out there not worth the money.

Thanks, I'm hoping for some input on this matter!

Welcome to Lymenet, birthdaysuit!

With that budget you should be able to get a pretty decent microscope. For doing digital captures it's very handy to have a trinocular. Adapters are microscope and camera specific, unless you get a universal one, like the one Lymedin2010 linked us to a while back.

Zeiss, Nikon, Olympus, American Optical, Reichert are good lab and research-grade scopes we recommend. Some here are doing excellent work with Amscope (like "dudefromkansas"), but he purchased a good 100x objective for his scope. The 100x objective his Amscope came with was useless.

You can see spirochetes in live blood with only 400x, but full 1000x is very handy.

We recommend a scope also equipped with darkfield, or phase contrast, or both. They make it very easy to see live objects without stain, but you can see live ketes with just brightfield, it's just more difficult.

To view Giemsa-stained slides all you have to have is brightfield, but you do need 1000x for that.

I bought my AO trinocular phase contrast scope for just over $100, but that was a really great deal.

Read over this whole thread, and you will get a better idea of what is involved with microscopy.

Here's another very informative forum I like:
http://www.microbehunter.com/microscopy-forum/

Thanks, TNT. In your opinion what is a good microscope for clarity and depth. The ones in my school are not able to see the spirochetes. I'm just looking for anything 1000 dollars and under that has decent zoom and clarity.
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birthdaysuit
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There's just so many options that it's overwhelming, that's all.
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birthdaysuit
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http://www.ebay.com/itm/OMAX-40X-2500X-USB3-10MP-Phase-Contrast-and-Darkfield-LED-Trinocular-Microscope-/400948263136?hash=item5d5a60f8e0:g:72sAAOSwLVZVksgr
_____
Is this decent?

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bluelyme
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The frame rate on those cams is iffy depending on the rig usb 3 may be ok ? But 10mp was shoddie ...
dude is using amscope dk 490 but added extra oil lens at 100x and is using his slr and is getting great videos..

inwould say olmpus nikon then american optical .but if you want new in box amscope over omax ..tnt has a ao, great rig see earlybon in thread and uses ccd sony cam

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birthdaysuit
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quote:
Originally posted by bluelyme:
The frame rate on those cams is iffy depending on the rig usb 3 may be ok ? But 10mp was shoddie ...
dude is using amscope dk 490 but added extra oil lens at 100x and is using his slr and is getting great videos..

inwould say olmpus nikon then american optical .but if you want new in box amscope over omax ..tnt has a ao, great rig see earlybon in thread and uses ccd sony cam

OLYMPUS microscopes are lovely but most of the OLY. Trinocular darkfield microscopes are over 1000 dollars.
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bluelyme
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Can someone tell me if this is a sting of pearls..and what the dancing artifact are (protozoan)?
Thanks
https://youtu.be/me-xa7ds8aU

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Blue

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TNT
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quote:
Originally posted by bluelyme:
Can someone tell me if this is a sting of pearls..and what the dancing artifact are (protozoan)?
Thanks
https://youtu.be/me-xa7ds8aU

It's a little hard to tell, blue, but, it appears to me to possibly be a kete with immune components (lysosomes) attacking it.

The dancing object does look a lot like an apicomplexan. I've seen identical flipping objects in my blood and I've seriously wondered about apicomplexans, but wondered how there could be so many at times.

Here are my videos on possible apicomplexans:

https://www.youtube.com/watch?v=nUZaE-NFlKw

https://www.youtube.com/watch?v=1eAGs5FzA2Y

https://www.youtube.com/watch?v=IcVMe35q7XE

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bluelyme
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Ya is that proto/toxo .? .your vids are amazing ! That biofilm in the last one and passing ketes to follow the apicomplexian ..just beautiful work tnt

does iver or babs tx hit those lil nasties?
How are you fairing these days ?

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TNT
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quote:
Originally posted by bluelyme:
Ya is that proto/toxo .? .your vids are amazing ! That biofilm in the last one and passing ketes to follow the apicomplexian ..just beautiful work tnt

Thanks for the compliments, Blue. I do not know if those objects even are apicomplexans.... but it sure does appear that way.


quote:
Originally posted by bluelyme:
does iver or babs tx hit those lil nasties?

Who knows what the treatment may be if they are some form of apicomplexan. I imagine Babesia treatment would help at least.



quote:
Originally posted by bluelyme:
How are you fairing these days ?

Not as good as I was, but I still believe the main issue is BLO, Mycoplasma, or a Rickettsia (or all three together).

The videos of possible apicomplexans I just linked to were from about a year ago. I haven't seen those in my last number of wet mounts.

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bluelyme
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Yes i think same here tnt ...i have a lot of the same structures in my stains as you posted above ..here is a quik vid of what looks like bart or rmsf ..like 70%of my cells are infected ?!

https://youtu.be/PrBdu0VyjpQ

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TNT
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Yeah, Blue, I'm really wondering if BLO or a Rickettsia is your main problem, too, especially after what happened to you on Bactrim.

Are you seeing the "Bart" dots only inside the perimeter of the RBC, or are you also seeing them on the outside of the cell wall of the RBC?

Just a FYI for those of you getting started with staining:

I don't put too much stock on dots being "Bart" unless I see them on the outside perimeter of the RBC and in an area on the slide that is clean of any other basophilic dots or artifacts. Some of my slides have had heavy presence of "dots" on the EDGES OF THE SMEARS, but I usually consider that contamination or artifacts.

For my own slides, I wish there was a way to eliminate any incidence & chance of contamination. I do my best to minimize that chance by using pre-cleaned slides and then washing my slides with soap and water before doing the smear. That way, any residual bacteria on the "pre-cleaned" slides is washed off. Also, I only use sterile pipettes to dip out of my stain. That way it's unlikely that I introduce any bacteria into my bottle of stain for later smears. I use distilled water to rinse the stain off the slides, and I try to make sure I wash my (reusable) gloves at the end. Even with these steps I wonder how many bacteria are contaminates, especially the basophilic ones.

Most times the contaminates are pretty obvious by changing focus in or out. If the bacteria are not on the same focal plane as the RBCs then it's pretty certain they are contaminates.


Blue, you're doing good work, so keep it coming. I'd be interested in seeing some more of those stained apicomplexans you showed us a while back if you come across any more. Though, it would be best that you don't, right!?

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bluelyme
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Thanks tnt for the much need encouragement ..i am inspiring some newbies on hw and it feels good visually see progress or not ...you are right this being the only cool part of this disease ..

here is that wbc and a short klip of blo/rmsf . On bottom rt of first one it looks pretty clear ?

https://youtu.be/rwD5tadvUZ0

https://youtu.be/P8kQm9Ddv4Q

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quote:
Originally posted by bluelyme:
Thanks tnt for the much need encouragement ..i am inspiring some newbies on hw and it feels good visually see progress or not ...you are right this being the only cool part of this disease ..

here is that wbc and a short klip of blo/rmsf . On bottom rt of first one it looks pretty clear ?

https://youtu.be/rwD5tadvUZ0

https://youtu.be/P8kQm9Ddv4Q

In that first video, that ring does appear to be Babesia, especially on account of the clearing in the center of the ring.

The second video, I would say they look more like Mycoplasma than BLO or RMSF. With Bart/BLO you usually see fewer organisms per field(than your second video showed), and the ones you do see are typically on the outside perimeter of the RBC.

Great job!

Yes, this is the ONLY cool thing about these diseases!

[ 11-16-2016, 07:29 PM: Message edited by: TNT ]

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Here is an EXCELLENT textbook article on Rickettsias!

(Todar's online textbook of bacteriology)-

http://textbookofbacteriology.net/Rickettsia.html

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TNT, where exactly is the anaplasma in your pictures? I'm not sure exactly where to look.

I have some stain coming in for the first time and my Igenex anaplasma came back with a hit, so interested to see if I can find some.

Edit: oh just realized these reside within the neutrophils? Never knew this!

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I've been watching Under Our Skin: Emergence at:

https://photos.google.com/share/AF1QipMPtByTwh0HHfjRgqmmu4c2U_uxoF-M_rWSRZT91wZENpHNfiK0kZN31iuXim8TqQ/photo/AF1QipOJjo5X7jra11NlhBB-pMuryrXfxpnQ4K91YzDz?key=WG5rcjU0aUMyODJIVV8zYW ZXR1pGS3dobXJUSFdn

About 47 minutes in, a Norwegian researcher says that he uses a thin solution of water and salt to see the organisms, this in a few minutes. He says that in principle you should be able to detect chronic borreliosis with a simple microscope

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quote:
Originally posted by mustardseed2:
TNT, where exactly is the anaplasma in your pictures? I'm not sure exactly where to look.

I have some stain coming in for the first time and my Igenex anaplasma came back with a hit, so interested to see if I can find some.

Edit: oh just realized these reside within the neutrophils? Never knew this!

Yes, in the pictures I posted of the neutrophils, they are the clump of purple organisms just inside the wall of the neutrophils.

The clump of elemental bodies inside the leukocyte is called a morula(e), which is a Latin word literally meaning (clump of) "mulberries."

You can also see what appears to be a morula in the picture of the platelet.

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quote:
Originally posted by mustardseed2:


Edit: oh just realized these reside within the neutrophils? Never knew this!

Anaplasma can also infect monocytes, platelets, RBCs, and be found free in the serum as well. I am pretty sure I am seeing some in biofilm, too.
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Hey bluelyme, I know you're doing rife therapy. Have you tried any of the Rickettsia frequencies? I am using Q Fever and it feels like it's knocking it back, or at least hitting it in a positive way.

Instead of using the typical "Bart" numbers, maybe try some of the other Rickettsia freq.'s.

Just a thought.

Have you done any recent slides and found any morulas? Are you still seeing many "Bart" dots on your most recent smear?

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quote:
Originally posted by lymenotlite:
I've been watching Under Our Skin: Emergence at:

https://photos.google.com/share/AF1QipMPtByTwh0HHfjRgqmmu4c2U_uxoF-M_rWSRZT91wZENpHNfiK0kZN31iuXim8TqQ/photo/AF1QipOJjo5X7jra11NlhBB-pMuryrXfxpnQ4K91YzDz?key=WG5rcjU0aUMyODJIVV8zYW ZXR1pGS3dobXJUSFdn

About 47 minutes in, a Norwegian researcher says that he uses a thin solution of water and salt to see the organisms, this in a few minutes. He says that in principle you should be able to detect chronic borreliosis with a simple microscope

INDEED!! A very simple microscope is all that would be needed! Even 400x can see spirochetes, although 400x in brightfield does not make it easy to see them. And, depending on how heavily infected you are, you probably would not even need the salt solution. I don't. Most chronic patients have many spirochetes, and those who are debilitated are loaded!
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I am looking at my stains and finding what appear to be diplococci within many of the leukocytes. So, I was wondering if these could be the Anaplasma parasite.

What I am finding is that, yes, they can indeed appear in that form, particularly during cell division (presumably before a complete morula develops).

Here is an excerpt from a veterinary book that goes into brief detail about the Anaplasma bacteria and contains pertinent illustrations.
"The Genus Anaplasma" starts on page 465.

https://books.google.com/books?id=SOVAAAAAIAAJ&pg=PA466&lpg=PA466&dq

[ 12-02-2016, 10:01 PM: Message edited by: TNT ]

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Hey tnt ..just got dna freq for bacciliformis ..i wil try q fever too...i just did aome smears of my folks ..step dad has erlichia and he had few wbc with mulberry action as you describe ...rifampin is kicking something as is houttynia ...maybe myco too i will post those smears soon .saw a weird inclusion almost as big as rbc all throughout his smear ?

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https://youtu.be/5-VABDNfDx0

https://youtu.be/qNx-BY5b65o

https://youtu.be/1fJQlXMVOgs

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Blue

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quote:
Originally posted by bluelyme:
Hey tnt ..just got dna freq for bacciliformis ..i wil try q fever too...i just did aome smears of my folks ..step dad has erlichia and he had few wbc with mulberry action as you describe ...rifampin is kicking something as is houttynia ...maybe myco too i will post those smears soon .saw a weird inclusion almost as big as rbc all throughout his smear ?

https://youtu.be/5-VABDNfDx0

https://youtu.be/qNx-BY5b65o

https://youtu.be/1fJQlXMVOgs


Great job, blue!

Yeah, those small, round, white clearings in the RBCs in that first video are weird. I'm not sure what to say about them. They definitely are "mystery inclusions." Unless you are referring to the "ballooned" edges of those 3 red blood cells. In that case, those could possibly be a result of oxidant injury. See Heinz bodies about halfway down this page:

http://www.eclinpath.com/hematology/morphologic-features/red-blood-cells/quick-guide/

Is the second video of the "possible erhlichia" what you are referring to in your step-dad's blood? IOW, is the second video your step-dad's blood? I would love to see the "weird inclusion almost as big as rbc."

Your third video, "Mycoplasma?," does look like Mycoplasma! It definitely is not contamination since it is only on the RBCs. The only other thing I can think it could possibly be is what is referred to as "stippling." Stippling frequently shows in chronically ill patient's blood, too. I'm not sure how one would tell the difference.

https://www.ncbi.nlm.nih.gov/pubmed/6202140

Here is a reference that may help show the difference:

https://quizlet.com/115897777/rbc-inclusions-and-infectious-agents-flash-cards/

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Here is a video about canine Erhlichiosis that popped up when I looked at blue's "possible Erhlichia" video. I'm not sure what the plug for the book at the end had to do with Erhlichiosis!

https://www.youtube.com/watch?v=v-Y00QYnM9Y

What stood out to me in the text was "no seasonality," "cerebellar ataxia," "paresis," "weight loss," "petechiae," "hyperplasia," and "vasculitis."

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Dr. H states the possible success of the Dapsone combinations may be because of a connection with Rickettsia bacteria:

https://www.facebook.com/drrichardhorowitz/posts/1153845914704205


Back in the 1970's when Willy Burgdorfer was sent blood from the Old Lyme patients to investigate what could be making them ill, he initially thought the causative agent to be a Rickettsia as he was finding this organism in many of their samples. But, after finding the spirochete, the Rickettsias took a back seat so to speak and this finding never really became known. Could this have been a factor that determined who became sick with Lyme disease in the first place, or perhaps who became chronic???

Obviously, Burgdorfer himself considered the Rickettsias a big deal--check out this article:

https://www.statnews.com/2016/10/12/swiss-agent-lyme-disease-mystery/


SPECIAL REPORT
The ‘Swiss Agent’: Long-forgotten research unearths new mystery about Lyme disease
By CHARLES PILLER @cpiller

OCTOBER 12, 2016

 -
- KRIS NEWBY/BURGDORFER ARCHIVES
A page from Willy Burgdorfer's archive shows elements of the research process he used to find infectious agents and study their properties. (English translation of the German: “Different Working Branches of Rocky Mountain Laboratories” )


The tick hunter was hopeful he had found the cause of the disabling illness, recently named Lyme disease, that was spreading anxiety through leafy communities east of New York City. At a government lab in Montana, Willy Burgdorfer typed a letter to a colleague, reporting that blood from Lyme patients showed “very strong reactions” on a test for an obscure, tick-borne bacterium. He called it the “Swiss Agent.”

But further studies raised doubts about whether he had the right culprit, and 18 months later, in 1981, Burgdorfer instead pinned Lyme on another microbe. The Swiss Agent test results were forgotten.

Now STAT has obtained those documents, including some discovered in boxes of Burgdorfer’s personal papers found in his garage after his death in 2014. The papers — including letters to collaborators, lab records, and blood test results — indicate that the Swiss Agent was infecting people in Connecticut and Long Island in the late 1970s.

And scientists who worked with Burgdorfer, and reviewed key portions of the documents at STAT’s request, said the bacteria might still be sickening an unknown number of Americans today.

While the evidence is hardly conclusive, patients and doctors might be mistaking under-the-radar Swiss Agent infections for Lyme, the infectious disease specialists said. Or the bacteria could be co-infecting some Lyme patients, exacerbating symptoms and complicating their treatment — and even stoking a bitter debate about whether Lyme often becomes a persistent and serious illness.

Swiss Agent, now called Rickettsia helvetica, is likely not a major health risk in the United States, in part because such bacteria typically respond to antibiotics. Still, several of Burgdorfer’s former colleagues called for infectious disease researchers to mount a search for the bacterium.


 -
-ALEX HOGAN/STATAfter initial tests, Burgdorfer suspected the Swiss Agent caused Lyme. He shared the strong evidence with a close colleague in Switzerland to see whether he could verify the findings in patients there.


“It should be done,” said Jorge Benach, a professor emeritus at Stony Brook University and a coauthor of Burgdorfer’s seminal 1982 paper describing the detection of the Lyme microbe. Public health concerns warrant a new study, Benach said, and with today’s more advanced “weaponry for pathogen discovery, it would make perfect sense.”

Dr. Paul Mead, chief of epidemiology and surveillance for the Centers for Disease Control and Prevention’s Lyme disease program, said that he wasn’t familiar with Rickettsia helvetica, but that “new tick-borne pathogens could certainly be out there.” He cited several found in the years since Lyme’s cause was discovered. Any serious, common co-infection would usually, but not always, be noticed by physicians as a distinct problem in Lyme endemic areas, he said.

In Europe and Asia, Rickettsia helvetica has been recognized as a relatively rare but sometimes serious health threat if untreated. It’s been linked to a handful of sudden deaths from heart disease, as well as facial palsy, deafness, meningitis, chronic muscle weakness, and temporary paralysis. But US laboratories don’t test for the Swiss Agent.

STAT was approached with Burgdorfer’s archives by Kris Newby, who is writing a biography of Burgdorfer and produced an award-winning documentary that sympathetically depicts Lyme patients and doctors who challenged the medical establishment over its approach to Lyme diagnosis and treatment.

The documents offer a tantalizing glimpse into how disease detectives tracked down Lyme’s cause — and how potentially significant loose ends can sometimes be dropped by researchers pressed for time and funding or diverted by more promising leads.

Burgdorfer note
BURGDORFER ARCHIVES
Note written by Burgdorfer
They show that Burgdorfer intended to look more deeply into the Swiss Agent, which he had discovered in 1978 in Switzerland, but never did. His former colleagues speculate that he set aside this research to focus on identifying the cause of Lyme. When the Swiss Agent turned out to be an unlikely candidate after all, he redeployed his limited time and resources to other prospects.

But the papers suggest that he might have gone to his grave harboring regret that he didn’t follow up on the Swiss Agent findings, as reasonable as the decision was, Benach said.

On the top of a stack of documents in his garage was a mysterious note, penned boldly in red ink in the scientist’s unmistakable handwriting. “I wondered why somebody didn’t do something,” it said. “Then I realized that I am somebody.”

The Lyme wars

Lyme has now become one of the most common infectious diseases in the United States — it’s been found in every state except Hawaii, and is rampant in the Northeast and parts of the Midwest. The CDC estimates that 329,000 people are infected annually.

Lyme has also provoked what’s often described as a “war” over diagnosis and treatment. If Rickettsia helvetica is in the United States, some experts consulted by STAT said, unrecognized infections might be one of several factors contributing to the controversy, by creating confusion over the cause of some patients’ illnesses.

The Infectious Diseases Society of America, the CDC, and many doctors view Lyme as generally easy to diagnose with its characteristic “bulls-eye” rash and pinpoint lab tests, and easy to cure with two-to-four weeks of antibiotics. If the disease is not diagnosed and treated early — in up to 30 percent of cases, there is no rash — patients can develop longer-lasting and more serious symptoms. But most infectious disease doctors say a short course of antibiotics will cure those patients.

But an insurgency of renegade doctors and patients disagrees. They argue that the diagnosis is frequently missed because of poor lab tests and other factors, and that Lyme becomes a chronic condition when untreated or inadequately treated. The patients describe symptoms that include incapacitating “brain fog” and weakness, intense anxiety, severe muscle pain, and paralyzing headaches. Many say that they required treatment with antibiotics lasting months or longer to be cured after years of misery.

Although the few small clinical trials that have examined long-term antibiotic therapy up to 90 days have shown few if any clear benefits, this camp has gained a passionate following, including a cadre of researchers who publish papers supporting this alternative view, and a medical group — the International Lyme and Associated Diseases Society.

The medical establishment mostly views “chronic Lyme” as the product of quack doctors exploiting desperate patients by offering unproven therapies. The patients sometimes need psychiatric care, these experts say, but in any case, chronic physical complaints are not caused by an active Lyme infection. Some state medical boards have gone so far as to revoke licenses of doctors who prescribe long-term antibiotics.

 -
-ALEX HOGAN/STAT
Dr. Allen Steere, then a professor at Yale who first identified Lyme disease in patients, was excited about initial lab tests that strongly suggested that the Swiss Agent was Lyme’s cause.

 -
- BURGDORFER ARCHIVES Burgdorfer in his lab.


It’s hard to overstate the animosity that characterizes this clash. A few angry patients have compared establishment Lyme experts — including Dr. Allen Steere, who collaborated with Burgdorfer and has received death threats — to the Nazi doctor Joseph Mengele.

How might the Swiss Agent add fuel to this conflict? Steere, a Massachusetts General Hospital researcher and among the world’s leading Lyme experts, said some patients who believe they have Lyme, but who test negative for the infection, might be suffering from an illness caused by one of several other microbes. Rickettsia helvetica could be among them, he said.

Ticks often carry more than one pathogen, so patients can also have co-infections along with Lyme, which frequently begin with similar symptoms, such as fever, neck stiffness, and headaches.

“You can’t tell them apart clinically” in the first several weeks, Steere said. Co-infections can cause “more severe early disease … a phenomenon of the summer, when the tick bites.” Longer term, the confusion would not last because of Lyme’s distinct symptoms, even if the infection were untreated, he added.

Other experts noted that Lyme and Rickettsia helvetica have co-infected patients in Europe. Antibiotics normally cure Rickettsia helvetica infections, but diagnosis can prove difficult because the microbe does not cause a rash. If untreated or inadequately treated, the two infections share overlapping, serious, and sometimes persistent symptoms, according to clinical researchers. These include debilitating fatigue, severe headaches, muscle weakness, meningitis, facial paralysis, and sarcoidosis — a chronic inflammatory disease that can cause lung and skin problems. Numerous studies have linked Rickettsia helvetica to such ailments, although it is not regarded as a major public health peril in Europe.

Andrew Main, who conducted Lyme research at Yale University in collaboration with Steere and Burgdorfer, had Lyme early on, before its cause was discovered, and was among patients who showed evidence of co-infection with the Swiss Agent — a result that was included in Burgdorfer’s papers but that Main knew nothing about until informed by STAT. The positive tests for the Swiss Agent among Lyme patients back then, he said, strongly support the idea that it might be a current threat.

Robert Lane, a University of California, Berkeley, medical entomologist and Lyme expert who worked closely with Burgdorfer, is respected by both sides in the Lyme wars. He said Rickettsia helvetica could be a significant hidden factor that worsens Lyme infections and makes them harder to cure.

“You would want to look at it both ways. Could that organism, if present in some of the Lyme-disease endemic areas, infect people and cause clinical illness on its own, or react in concert with (the microbe that causes Lyme) or some of the other agents,” Lane said. “If you are looking for one or a few agents in a tick, you may be overlooking others that contribute to the disease burden.”
Finding the Swiss Agent

The man who found Lyme’s cause devoted his career to studying creatures sometimes described as tiny living cesspools, for the infectious stew of microbes ticks carry and transmit while sucking blood from animals or people.

While training for his PhD in his native Basel, Switzerland, Burgdorfer became a preeminent “tick surgeon,” as he called himself — dissecting thousands with eye scalpels and Swiss watchmaker forceps. In 1951 he became a research fellow at the federal Rocky Mountain Laboratories, a remote outpost in Montana’s breathtaking Bitterroot Valley that specializes in infectious agents.

Burgdorfer fell in love with the Bitterroot and with Gertrude Dale See — a secretary and technician at the lab. She won the multilingual scientist’s heart with her ability to speak French. They married and had two sons, and Burgdorfer became a US citizen and permanent lab employee.

He rose to lead the work on Rickettsia, rod-shaped bacteria spread by ticks that cause ailments such as Rocky Mountain Spotted Fever — which is sometimes deadly for patients in New England as well as the West. Burgdorfer built a global reputation for his knowledge of Rickettsia and Borrelia — corkscrew-shaped “spirochete” bacteria of the same group as the species known for causing syphilis.

On a trip back to Switzerland in 1978, Burgdorfer and a few colleagues discovered in local ticks the previously unknown Swiss Agent — later named Rickettsia helvetica (from Switzerland’s ancient Latin name, Helvetia). He found the microbe infectious for meadow voles — a small rodent common in Europe and the United States — and deadly to chicken embryos. No one knew then that it also caused illnesses in people.

Burgdorfer returned with samples of infected ticks and Swiss Agent antigen, molecules from the bacterium that can provoke an immune response, for further study. When mixed with blood sera — a part of the blood that doesn’t contain blood cells — the antigen can show whether a person has been infected.

By then, Steere, a young Yale professor, had for several years been aggressively investigating why some of his patients in Lyme, Conn., were reporting serious and strange symptoms of an apparently new illness. He had found “that many patients suffered not only of arthritis, but also of disorders affecting the skin, muscular, cardiac, and nervous systems,” Burgdorfer told his official biographer from the National Institutes of Health in 2001.

Steere asked Burgdorfer to join the hunt for a tick-borne microbe believed to be at the heart of Lyme. He sent samples of his patients’ blood sera to Rocky Mountain Laboratories for analysis.


 -
-ALEX HOGAN/STATBlood sera from Lyme patients showed infection with the Swiss Agent. Results of 64 or greater were considered firm evidence, as this test showed for 6 of 11 patients. The test showed no infections with other Rickettsia.


Sera tests showed that at least a dozen Lyme patients had been infected with Swiss Agent, and that at least six others might have been infected. The records did not make clear how many Lyme patients had been tested overall. Burgdorfer told Steere and other colleagues that the results pointed to a potential cause of Lyme.

Steere sensed a breakthrough. “I am excited to pursue further the possibility of a rickettsial etiology of Lyme disease,” he wrote to another researcher.

Burgdorfer was encouraged, in part, because of the test’s specificity: A positive result strongly suggested that the person had been infected with the Swiss Agent and not a different Rickettsia such as the one that causes Rocky Mountain Spotted Fever.

But when a second test method showed inconsistencies, doubts crept in about whether Swiss Agent was linked to Lyme. About 18 months later, Burgdorfer broke through, providing a rare undisputed fact in what would become the most disputatious of diseases: A spirochete causes Lyme. Years later, the microbe was named in his honor, Borrelia burgdorferi.

But he hadn’t given up on Swiss Agent completely.

In the lab during this period, Burgdorfer infected US ticks with the Swiss Agent, his lab books show. The records don’t state his experimental goal, but Rocky Mountain Lab scientists often studied which animals and arthropods could be infected with different agents, and thus might be reservoirs or vectors for disease. He also looked for Rickettsia in ticks in Lyme-endemic areas and found dozens of examples, but often neglected to determine the specific rickettsial species.

In December 1981, just a few months after discovering the Lyme spirochete, he wrote to a Swiss colleague who was overseeing a young investigator’s defense of his PhD thesis concerning the Swiss Agent. Burgdorfer suggested this question: “Do you feel that ‘Rickettsia suisse’ is the etiologic agent of (Lyme)? If so, how would you go about proving this?”

Burgdorfer and his colleagues reported their discovery of the cause of Lyme in the journal Science in 1982. In a handwritten draft found among Burgdorfer’s papers, he described identifying Rickettsia in Lyme patients’ sera and ticks, and his efforts to rule out Rickettsia as the cause of Lyme — without naming the Swiss Agent.

But in the final Science article, he made no mention of Rickettsia. Not a word about possibly finding the Swiss Agent in this country has ever been published.

Finishing the hunt

Burgdorfer retired in 1986 at age 60, just a few years after the successful Lyme hunt put him at the pinnacle of his field.

“I started to realize that the research I used to do and was successful in doing has changed its character,” he explained to a National Institutes of Health biographer in 2001. “Molecular and genetic biology have replaced the technologies I was able to apply,” he said. “Since I had no basic training in these fields … I was unable to speak and understand the completely new language.”

Those fluent in the “new language” of molecular biology and genetics will be able to finish Burgdorfer’s work, experts said. If the Swiss Agent is here, they can find it.

The CDC’s Mead said his agency is using molecular techniques to look for evidence of bacteria in 30,000 sera samples from people suspected to have contracted tick-borne illnesses. If Rickettsia helvetica is in some of the samples, it probably will be found, he said. That process will taken several more years to complete.

Dr. W. Ian Lipkin, who directs the Center for Infection and Immunity at Columbia University, is hunting for viruses as well as bacteria living in ticks that spread Lyme, partly to understand why antibiotics sometimes fail in apparent Lyme cases.

Lipkin’s group has collected 5,000 ticks from New York and Connecticut. With funding from the Steven and Alexandra Cohen Foundation, he has so far identified 20 new viruses in these ticks, and is exploring whether they have caused harmful infections in people, using tests that can search for a wide range of tick pathogens in a single sera sample. Eventually, Lipkin said, this process could make the tests affordable on a mass scale.

“Everyone wants to get to the bottom of this,” Lipkin said. “All of this is critical to … finding out why some people respond to antibiotics and some people don’t, and whether or not the antibiotics being used are appropriate, and trying to find ways to link different bacteria and different viruses to different syndromes.”

Lipkin is seeking funds to expand the work to tick-borne bacteria, including Rickettsia.

Asked whether his methods could find evidence of infections with the Swiss Agent, Lipkin replied without hesitation. “The answer is yes,” he said. “If this particular rickettsial species is present, I’m sure we will see it.”


 -
-KRIS NEWBY/NATIONAL ARCHIVES AND RECORDS ADMINISTRATION ARCHIVESNegatives of microscopic images of the Swiss Agent, from Burgdorfer’s archive


Willy’s last words

After he retired, Burgdorfer sent most of his voluminous personal files to the National Archives in Washington, D.C., where they were cataloged for public viewing. Those records contained some Swiss Agent documents. Many more lay untouched for decades in his garage and home office in Hamilton, Mont.

Late in life, Burgdorfer developed Parkinson’s disease and became increasingly infirm. A friend listened to his fears that his garage files might be lost to history. She urged Burgdorfer to contact Ron Lindorf, then an entrepreneur and business professor at Brigham Young University, who had been suggested by colleagues.

Early one morning in June 2014, an agitated Burgdorfer called Lindorf with an urgent request: “Come to Montana and get all my research, my files. I want to put it on the internet so people can see it,” Lindorf recalled him saying.

Lindorf was not a professional archivist, but agreed: His children had suffered from serious bouts of Lyme disease, he was eager to help the scientist who discovered Lyme’s cause, and he had the ability to take on the complex job. The next month Lindorf arrived in Hamilton, departing two days later with his SUV packed full of old files. That November, Burgdorfer died.

To better understand the Burgdorfer archive, Lindorf began collaborating with Newby, producer of “Under Our Skin,” the Lyme documentary. She shared the documents with STAT, hoping that an independent report would illuminate a possibly hidden risk for Lyme patients and others.

Lindorf returned to Montana last year to visit Burgdorfer’s second wife. She pointed across the garage to some additional boxes. Inside a cardboard portfolio covered in flowery fabric and closed by a metal clasp, he found more of the Swiss Agent archives, topped by Burgdorfer’s “I wondered why somebody didn’t do something” note.

“It made the hairs on the back of my neck stick up,” Lindorf said. “It felt like Willy talking from the grave.”


_______________________________________

Charles Piller can be reached at [email protected]
Follow Charles on Twitter @cpiller

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bluelyme
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Thanks tnt ..great info re swiss agent .cant wait for uos to release doc ....i got the script for dap but its so close to a sulfa i am scared stiff after bactrim episode ..the vasculitis is sorta calmed down after much iv curcumin and rocephin

--------------------
Blue

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TNT
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quote:
Originally posted by bluelyme:
...rifampin is kicking something as is houttynia ...

I love Houttuynia! And CSA! Both have really helped me over the past year. Not curative by themselves....but very synergistic with ABX.

Just had to put my plug in about herbals. Wonderful medicine. And Stephen Buhner is GENIUS!

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thatdudefromkansas
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I'm back to get involved for a bit.

Been traveling and had some other things come up.

I am gonna prepare a batch of the BSK-H without the antibiotic mixture.


If anybody needs some, just message me. I might be able to help out.

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Lymedin2010
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Staphylococcus aureus, the one that causes biofilm & issues with joint replacement prosthetics & any other foreign objects in the human body (this is well known). Also the cause of skin infections & respiratory infections, including MRSA.

I would imagine this may be an added issue for Lymies & I never hear of anyone talking about this. I can see this not only being in a Lymie body, but living within Borrelia biofilm.

https://www.youtube.com/watch?v=1xlNgeBc3ek

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Lymedin2010
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CytoViva is a huge microscope & imaging manufacturer.

"Rickettsia Bacteria Intracellular: Captured Using CytoViva Enhanced Darkfield Microscopy System "

https://www.youtube.com/watch?v=cv_7I8D4KQc

______________________________________

Spirochetes in Live Blood Cell Culture: Captured Using CytoViva Enhanced Darkfield Microscopy
https://www.youtube.com/watch?v=DnsuiSKGDRs

______________________________________
Nice images of candida.
http://msphere.asm.org/content/1/6/e00187-16

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TNT
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Amazing!

Lymedin, you always amaze me with what you uncover! That CytoViva video of the Rickettsia is exactly what I am seeing in my live blood from the beginning! Remember the "Bartonella" organism I showed video footage of? We debated about whether they were dumbbell ketes, or a Bart-like bacteria. I finally decided it must have been overflow from leaky gut. Now I'm sure they are Rickettsias! Probably Anaplasma. Especially since I'm finding unmistakable morulas in my stains. It would be interesting to know what species of Rickettsia the CytoViva video is showing.

Here are the two videos I had posted that document my Rickettsia bacteria:

https://www.youtube.com/watch?v=lOftWSU92fY

https://www.youtube.com/watch?v=F331jqs3qaE

Lymedin, thanks for that video, and of the one of Borrelia. It's so good to have lab verification of carbon copies of what we are seeing under our own scopes!


dudefromkansas, welcome back! I'm sorry to hear about the scope. FWIW, there is a well-equipped Zeiss Photomicroscope III on Ebay right now for about 3 grand. I don't know what your plans are or what you may now be looking for, but it's a sweet scope:

http://www.ebay.com/itm/CARL-ZEISS-PHOTOMICROSCOPE-MICROSCOPE-ACHR-APL-1-4-0-32-CONDENSER/231566477918?_trksid=p2047675.c100005.m1851&_trkparms=aid%3D222007%26algo%3DSIC.MBE%26ao%3 D2%26asc%3D38530%26meid%3D4568cafd94fc49bba5db59b419440f3d%26pid%3D100005%26rk%3D1%26rkt%3D6%26sd%3D261893232493

And, another:

http://www.ebay.com/itm/Carl-Zeiss-PhotoMicroscope-III/131735280345?_trksid=p2047675.c100005.m1851&_trkparms=aid%3D222007%26algo%3DSIC.MBE%26ao%3D2%26asc%3D38530%26meid%3D206d75ada 3b341c8b6301d54a5db2b74%26pid%3D100005%26rk%3D4%26rkt%3D6%26sd%3D331753091995

I hope you are doing well physically, dude. Keep up the great work!

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thatdudefromkansas
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Yes, I am shopping for a scope.

Just gotta make sure it's right for what I need, and I am also now wary of buying used due to the last experience.


I have contemplated starting with a lower end fluorescent capable microscope.

However, I also want to enhance my culture methods first. So my work now will be focused on improving the culture method before I think about doing anything more advanced than what I am doing now.

It's time and money, so gotta make sure it is done right.

Great videos and great research provided everyone.
Maybe we could get some sort of collaboration going on some of this stuff, direct collaboration.

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bluelyme
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tnt that second vid was awesome , that blo was chasing the kete with a vengance , i hope they werent colaborating but if we could just sick em on the ketes ha ha , or is phage therapy any kind of real probability with regards to risk vs benifit

was like watching a mantis fight

tnt posted https://www.youtube.com/watch?v=F331jqs3qaE

--------------------
Blue

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thatdudefromkansas
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TNT, I want to say you are doing some good work with your stuff, and it has motivated me a bit to begin my work again.

Excellent

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TNT
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quote:
Originally posted by thatdudefromkansas:
TNT, I want to say you are doing some good work with your stuff, and it has motivated me a bit to begin my work again.

Excellent

Hey, thanks! I really appreciate the compliment! But, I have to say, you guys have been a great inspiration to me! It was this thread that first showed me what all was possible with microscopy. The work of Lymedin2010, S13, and PeterKemp was a huge motivation for me to get involved, especially Lymedin's. It's hard not to catch Lymedin's enthusiasm.

And, dude, your pioneering & culturing skills are really amazing! To come up with homemade BSK medium is genius! And your video capture of the kete to cyst conversion was just timeless and priceless! I think it's great we have slightly differing interests among us. It's very complimentary.

I've learned a lot, and much of it is because of you guys here! Keep up the good work fellows! And that's to all of you-- those of you who are veterans, and those of you who joined more recently.

I hope you new guys become regular contributors and this really becomes a movement.

I also wish S13 and PeterKemp would come back on and give us their great input! They both have some great microscopy skills, not to mention Lymedin2010.

THE MICROSCOPY THREAD ROCKS!!!

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Lymedin2010
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Peter Kemp is on FaceBook & is active in the LD community. We have all hit a limitation on what we can do with a microscope, other than to see the spiros.


I have contacted Cytoviva on that video about Borrelia in blood & this is what they had to say...


"Thank you for your interest in CytoViva technology and the ability to observe Borrelia and related organisms. Our patented enhanced darkfield microscopy enables the capture very high signal-to-noise images of these types of pathogens in-situ in tissue, blood cells and other environments.



By adding hyperspectral imaging to the microscope, you can spectrally characterize these pathogens in these different environments.



The video you referenced was captured by microbiology researchers at Auburn University who were studying Borrelia. These bacteria were specifically cultured by this microbiology group.



CytoViva provides fully equipped optical microscopes for this type of imaging and can also provide integrated hyperspectral imaging as required.



Please let us know how we can support you with more insight regarding this technology.



Kind regards,"

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thatdudefromkansas
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I'm hoping to convince my neuro for a CSF sample on my next appointment.

It would be great to analyze, culture, and stain that.

If I got a CSF sample, I would definitely ensure that I invest in adequate fluorescence microscope and stains.

We'll see how that goes.

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thatdudefromkansas
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Any idea on the cost of those CytoViva microscopes.

I'm guessing in the range of 30,000 dollars.

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Lymedin2010
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TNT, I have seen what I called "bean' like objects in my blood too & it looks very similar to Ricketssia. I always thought they were the discoid gemma of the spiros. We do have one capture of a spiro that morphs into this structure & I would have to dig it up again. In tick juice, I do not see any discoid gemmas, but I have seen fairly large cysts & cyst small enough that they get burried & are indistinguishable from the tick gut cells.


Yea, I too would love to obtain a CSF sample. I had done 2 spinal taps before I had a microscope & have not had one since.

I am so tempted to extract some sinovial fluid from my knee at one point, as I am expecting to see more spiraling forms in that fluid.

None of the Cytoviva prices are listed, so they must be in the thousands. No need for a Cytoviva scope, as the new 4K addition the any scope will bestow it with grand improvements. Look into a good 4K camera with an adapter, such as the Canon SLR's or Sony Alpha cameras.

Check out these good fluorescent scopes you can put a lower bid amount.

http://www.ebay.com/itm/Nikon-Microscope-Eclipse-E200-with-Fluorescence-/162293158902?hash=item25c96d13f6:g:KHQAAOSwSWJXRLNq


http://www.ebay.com/itm/Nikon-Labophot-2-Fluorescence-Microscope-/182384796291?hash=item2a76fb2283:g:cvMAAOSwHMJYMJXG

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TNT
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quote:
Originally posted by Lymedin2010:
TNT, I have seen what I called "bean' like objects in my blood too & it looks very similar to Ricketssia. I always thought they were the discoid gemma of the spiros. We do have one capture of a spiro that morphs into this structure & I would have to dig it up again.

Yes, definitely dig that one up, I would love to see it! I have seen the disc-shaped cysts of borrelia with my darkfield (like what dude showed us in the conversion video) and there is no mistaking the cysts. Darkfield seems to be the best technique for viewing them.

Also, were you able to find those pics of the other lyme patient's morulas? I assume they were on Facebook? I would enjoy seeing them, too!

quote:
Originally posted by Lymedin2010:
We have all hit a limitation on what we can do with a microscope, other than to see the spiros.

Yeah, I can relate to an extent. I'm just encouraging all of us to post the things we do see, even if similar things have already been posted. If nothing else, it keeps the thread alive and keeps our interests up.

I'm a little unclear if you have done any Giemsa smears, Lymedin. It's an indispensable tool for particular infections, as you already know. Also, I was wondering how you are doing these days. Since we haven't heard much out of you lately, I figured you were still pretty low. That's been my impression. Maybe you are doing well enough you are busier with life. What are you doing for treatment now? Have you considered BVT??? I am making some real progress on it, along with herbs and an ABX or two. The BVT didn't really start kicking butt until I got to the nominal dose, which is 10 stings per time. But, it took time to build up to that. Of course, maybe you've already considered it since you follow Eva Sapi's work very closely.

And, S13, if you are still out there, pop in and give us an update and show us anything of interest! Anything.

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TNT
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quote:
Originally posted by Lymedin2010:
TNT, I have seen what I called "bean' like objects in my blood too & it looks very similar to Ricketssia. I always thought they were the discoid gemma of the spiros. We do have one capture of a spiro that morphs into this structure & I would have to dig it up again.



quote:
Originally posted by TNT:
I have seen the disc-shaped cysts of borrelia with my darkfield (like what dude showed us in the conversion video) and there is no mistaking the cysts. Darkfield seems to be the best technique for viewing them.

In the same darkfield sample as the cysts, I also clearly saw what you are calling "bean-shaped" organisms. So, I am pretty sure what I am seeing are not the same things. I'll go back and look at that and maybe put it up.

[ 12-13-2016, 02:00 PM: Message edited by: TNT ]

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TNT
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Lida Mattman's 4th edition of Cell Wall Deficient Forms- Stealth Pathogens is finally being released and you can order your copy now!

It is on a pre-order sale at Amazon for $149.95!

I just thought I would let you guys know!

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thatdudefromkansas
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Anybody have any luck with increasing resolution with new, higher grade objectives?

Like switching out for Zeiss or Leica or anything?

I am curious how much difference it would make on a lower end microscope like my Amscope.

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thatdudefromkansas
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Anybody have any luck with increasing resolution with new, higher grade objectives?

Like switching out for Zeiss or Leica or anything?

I am curious how much difference it would make on a lower end microscope like my Amscope.

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thatdudefromkansas
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Lymedin, Staph. Aureus is an extremely common bacteria that can cause significant infections. Usually a dermatological infection, anytime someone says they got staph, that's it. Cuts, abrasions, etc, that get infected are generally always caused by staph aureus.
It's a common infection in surgeries, and is always treated prophylactically for that.

It's also a concern in our microscopy, especially live microscopy, because it is one of the most common contaminants.
It can render cultures useless.
If you touch something ungloved, you might contaminate it.

If you want a break from staining for Borrelia, take some swabs of your skin. Practice and refine staining that. You'll get staph. aureus on a skin swab. You can even just take a sterile q tip and dab a bit of saline on it, swab your skin, and then transfer that to a slide. It's a good way to practice. And you can get practice differentiating between the different types of bacteria and determine gram negative or gram positive, etc.

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Lymedin2010
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TNT, thanks for asking. I've been trapped in hell, as we all have, with many pains, exhaustion & my brain is on fire & hence less posts on here.


We have all become more experienced here & it requires a bit more vigor & brain power. Every now & then I got a slight relief from the brain fog. ABX & even herbs seem to have just stopped working.


I had done Giemsa in 2012 & I bought a cheaper batch and the stains were filled with tons of artifacts. I have just purchased the same strain from your recommendations & could probably use a lesson in the finer details & any tricks of the trade that you might have happened to stumble upon.


Dude, yea I took a Courseca course on Aureus & it was very interesting, as I did not know these things before the course. Ever since that course I have been upon to tertiary infections having a profound impact on Lyme as a whole & probably from multiple pathogens more likely.

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Lymedin2010
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When we asked Alan MacDonald about the My Horrific Lyme Video in an email, this is what he responded within that email & he then posted his response publicly on LymenetEurope. You can check the post for yourself.


"Dear Zoran,

I have viewed your video and I concur that your images do indeed demonstrate Spirochetal forms

in various profiles. I would avoid the word "debris" because I have evidence from my DNA Probe studies

with Probes absolutely specific for Borrelia DNA ( two sites on The Chromosome- Burgdorferi and two sites on the

chromosome of Borrelia Miyamotoi) that each and every DNA containing membrane bound Borrelia

variant ( spirals, cylindricals, Segmented, String of Pearls, Granular forms, and Liposome (Blebs) , and Cystic forms

are ALL CAPABLE of regenerating the spiral Borrelia form under ideal circumstances. You have described

"medusa" profiles of Borrelia, and this is a good name for what you see in some of your images.

You have not described the Biofilm Borrelia communities. These are extremely important, and my work has proven with

DNA Probes for Borrelia DNA ( Molecular Beacons) the Biofilm communities of Borrelia

dwell Inside of Amyloid Plaques in Alzheimer's disease. I attach some images for you to inspect from DNA Probe hybridization

of Biofilm Borrelia Communities ,Inside of human Autopsy Brain, surrounded by Amyloid in Plaque formation.
" http://www.lymeneteurope.org/forum/viewtopic.php?f=7&t=5873&hilit=Research+funding+appeal+DrMacDonald+DNA+PROBES+DEVELOPMENT

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TNT
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quote:
Originally posted by Lymedin2010:
TNT, thanks for asking. I've been trapped in hell, as we all have, with many pains, exhaustion & my brain is on fire & hence less posts on here.

We have all become more experienced here & it requires a bit more vigor & brain power. Every now & then I got a slight relief from the brain fog. ABX & even herbs seem to have just stopped working.

I'm so sorry to hear this, although I have been suspecting it. I have been having success with BVT and I don't think you should try it.... I KNOW you should try it!! It's better than CBD! REALLY! I forget where I put my prescription pad, otherwise I would write you a script for it! [lol] [lol]


quote:
Originally posted by Lymedin2010:
I had done Giemsa in 2012 & I bought a cheaper batch and the stains were filled with tons of artifacts. I have just purchased the same strain from your recommendations & could probably use a lesson in the finer details & any tricks of the trade that you might have happened to stumble upon.

I think the Romanowsky stain will give you a clue as to what is going on. Unless it simply is Borrelia out of control, or something like a virus.

Did you get the "Blue" or the "Red" stain from that company? Either way, they are very similar and should be able to see the same things. The red is equivalent to straight Wright's stain, and the blue is equivalent to Wright-Giemsa. Both are Romanowsky, and either one will be very helpful.

There is no mixing with those products, so there really are no steps other than having a dried smear and staining it. After preparing my slide I allow the blood to dry by placing it in my oven at 100 degress F. That temp is used when doing a smear for Malaria so that the parasites don't destruct in the process of drying. I do it that way too in case it's the same for other apicomplexans.

I draw out of my bottle of stain very carefully with a sterile pipette. I have found that you get artifacts if you don't sufficiently flood the top of the slide. That said, you still only draw out maybe a 1/2 an ml, or less. So, only a very small amount is used. But, if you don't use enough stain, it will begin to dry on the slide before you rinse, and that produces the artifacts. After allowing the stain on the slide for 2-3 minutes, I drop the slide in a coplin jar of distilled water and allow to soak for about 1-2 minutes before rinsing with distilled water from one of those lab squirt bottles. Allow to dry in a covered container (to minimize airborne contamination), and you are ready to view.

I plan on posting some pics and video from my darkfield demonstrating the difference between cysts and Rickettsia-like bacteria. Stay tuned.

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Lymedin2010
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There is a special PCR DNA sequencing going on now at uBiome.

Sequence all 5 locals from: gut, nose, skin, mouth, & genitals for less than $100, rather than the $400 usual price.

I added my blood to the nose sample, as sometimes there is blood in the nose anyways. Many times there is blood in my nose, so a little more won't hurt. [Smile]


I'll let you guys know mine when it comes back.


https://www.facebook.com/uBiome/photos/a.138580672964326.30062.109232119232515/730027620486292/?type=3&theater


https://ubiome.com/explorer

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Lymedin2010
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I bought the blue one. Thanks for the instructions & it sounds easy enough, but it always does on paper & a bit more messier & tricky in person until you have done it a few times.
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