For those of you doing your own microscopy... *Part 3*

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bluelyme
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   Posted 9/11/2017 4:17 AM (GMT -6)   
/youtu.be/oWw4kV5GSv8 lil ketes .

hey does anybody know if on the amscope t490b - do i have to get a darkfield oil lens at 100xand a new condeser? and rig a light to use at that power

bluelyme
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   Posted 9/11/2017 4:34 AM (GMT -6)   
Also i am seeing a lot round things that spirochetes are attached or coming out of are the Lform? doe any body have links to lida mattman pics ?

ChickenArise
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   Posted 9/11/2017 9:31 AM (GMT -6)   
Healrom said...
Yes, I know him. He bought a fluorescence microscope. And he begins to make very good videos with it!


Is this the same as a UV light microscope? How close to becoming affordable are these UV light microscopes today?

I am not experienced in this area but it sounds like all kinds of once hidden organisms and processes are on the verge of being (re)discovered due to prices becoming more reasonable.

Are there any YouTubers using these and making videos today? I've reached a limit in what I can find online.
AUG14:Mold Sick.FALL16:Clinical Bart/Borellia/Yeast
NOV16:Lung Pain. JAN17:Morg Scalp (resolved)
FEB17: Pupils, throat glow UV light.
Rx: Abx Break, Probiotic Rebuild, FLZ q72hr
Sup: Ozone, Magnets, Dental implants removed
Proto:Modified Klinghardt
Tx: self

Mustard Seed
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Date Joined May 2016
Total Posts : 1087
   Posted 9/11/2017 1:11 PM (GMT -6)   
bluelyme said...
Also i am seeing a lot round things that spirochetes are attached or coming out of are the Lform? doe any body have links to lida mattman pics ?


I believe those round, wobbling things are cyst (round bodies). Watch the conversion in real time:

/m.youtube.com/watch?v=1HUtKungjvE

bluelyme
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   Posted 9/12/2017 12:00 PM (GMT -6)   
Great vid mustard ...wow ...any ideas @ darkfield at 1000x
maybe toots can explain flourencence microscopy a bit

Healrom
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Date Joined Nov 2016
Total Posts : 133
   Posted 9/12/2017 9:53 PM (GMT -6)   
Mustard Seed said...


I believe those round, wobbling things are cyst (round bodies). Watch the conversion in real time:

/m.youtube.com/watch?v=1HUtKungjvE


It's not in real time. Speed 8x. The woman that made the video used a mix with acridine orange. It's a fluorescent stain (but used in BF here).


ChickenArise said...

Is this the same as a UV light microscope? How close to becoming affordable are these UV light microscopes today?

I am not experienced in this area but it sounds like all kinds of once hidden organisms and processes are on the verge of being (re)discovered due to prices becoming more reasonable.

Are there any YouTubers using these and making videos today? I've reached a limit in what I can find online.


A fluorescence microscope uses lights that emits UV, yes. All the stains don't react to the UV wavelenght, but some yes. It depends. So you put the good light filter for the good stain, etc...
He bought a second hand microscope, but I don't know how much it cost him at the end. You should ask him on his channel, or on FB. His name is Jack Dupre Stravinsky.

Joyous
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   Posted 9/12/2017 10:22 PM (GMT -6)   
Healrom - are those spirochetes in your video that are wiggling???

Healrom
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Date Joined Nov 2016
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   Posted 9/12/2017 11:18 PM (GMT -6)   
In which one?

Mustard Seed
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Date Joined May 2016
Total Posts : 1087
   Posted 9/13/2017 10:48 AM (GMT -6)   
bluelyme said...
Great vid mustard ...wow ...any ideas @ darkfield at 1000x
maybe toots can explain flourencence microscopy a bit

If you're looking at the T490B and want to do 1000x darkfield, you'll need to buy a few things separate: an oil darkfield condenser (N.A of ~1.25), an 100x oil objective with an adjustable iris (or a funnel stop with the existing objective), and likely a brighter light source. In the Lymenet thread there was a guy who made this conversion, though I don't remember the name.

Also, what camera did you use to capture the video that you posted?

Healrom said...
It's not in real time. Speed 8x. The woman that made the video used a mix with acridine orange. It's a fluorescent stain (but used in BF here)

You're right about that, somehow I missed that. The brownian motion looked remarkably like real time but it's not.

Joyous said...
Healrom - are those spirochetes in your video that are wiggling???

If you're talking about the video blue posted, then yes those wiggling things are spirochetes. Same with the video I posted. The transformation into the cyst in the latter video makes me believe the wiggling round balls in blue's video are also cysts. I find the same things in my blood.

When I'm on antibiotics I find almost exclusively cysts until the sample has been left out for 24-48 hours, when the spirochetes come out to play. When I was on Flagyl, there were no cysts in my blood, but I found many spirochetes right away without having to let the sample sit for 48 hours.

ChickenArise
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Date Joined Nov 2015
Total Posts : 1052
   Posted 9/13/2017 3:31 PM (GMT -6)   
Healrom said...

ChickenArise said...

Is this the same as a UV light microscope? How close to becoming affordable are these UV light microscopes today?

I am not experienced in this area but it sounds like all kinds of once hidden organisms and processes are on the verge of being (re)discovered due to prices becoming more reasonable.

Are there any YouTubers using these and making videos today? I've reached a limit in what I can find online.


A fluorescence microscope uses lights that emits UV, yes. All the stains don't react to the UV wavelenght, but some yes. It depends. So you put the good light filter for the good stain, etc...
He bought a second hand microscope, but I don't know how much it cost him at the end. You should ask him on his channel, or on FB. His name is Jack Dupre Stravinsky.


Thank you for taking the time to educate a newbie on the subject and for the reference to someone using UV microscopes.

Grayfieldoptical UV microscopes claim you dont need a stain, but they dont put their prices on their site so I would imagine these things still cost a pretty penny although they are working their way down in price and eventually will reach the affordability of the hobbyist.

I know I am in the minority to even entertain the pleomorphic theories of old but it sure would explain why all these different species can communicate with each other in quorum sensing if they were all originating from a somatadid.

As the price drops in these microscopes if there is anything being hidden from us, they wont be able to hide it forever as the enthusiasts will expose what they witness, so until that time I will just keep an open mind and wait and see what develops.

Thanks again for educating me.
AUG14:Mold Sick.FALL16:Clinical Bart/Borellia/Yeast
NOV16:Lung Pain. JAN17:Morg Scalp (resolved)
FEB17: Pupils, throat glow UV light.
Rx: Abx Break, Probiotic Rebuild, FLZ q72hr
Sup: Ozone, Magnets, Dental implants removed
Proto:Modified Klinghardt
Tx: self

gfields
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Date Joined Oct 2015
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   Posted 9/13/2017 5:41 PM (GMT -6)   
Here's some good links on microscopy. I think I may get one. I've been procrastinating.


http://www.healingwell.com/community/default.aspx?f=30&m=3756602

http://www.healingwell.com/community/default.aspx?f=30&m=3760106

http://www.microscopenet.com/omax-40x2000x-darkfield-trinocular-compound-microscope-with-digital-camera-p-10243.html?gclid=CjwKEAiAvs7CBRC24rao6bGCoiASJABaCt5DpGWSgZWPThqIzpZd7NIpz8gYP9-mpvnmXC-6TS9pqRoCVrnw_wcB

Healrom
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Date Joined Nov 2016
Total Posts : 133
   Posted 9/13/2017 6:17 PM (GMT -6)   
Mustard Seed said...

When I'm on antibiotics I find almost exclusively cysts until the sample has been left out for 24-48 hours, when the spirochetes come out to play. When I was on Flagyl, there were no cysts in my blood, but I found many spirochetes right away without having to let the sample sit for 48 hours.


Yes, me too. Exactly the same. Also when I have cysts, sometimes I find thousands of thin spirochetes. (2X thinner than the normal ones)

It's for this reason that we have to combine multiple treatments.
The best combo that I have found until now is the buhner protocol : cat's claw and andrographis. CC for spirochetes and andro for cysts.
Also with the japanes knotweed, but it doesn't seems as effective.

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 224
   Posted 9/14/2017 2:04 PM (GMT -6)   
bluelyme said...
maybe toots can explain flourencence microscopy a bit


Typical vertical fluorescent microscope setup:

VIEW IMAGE

I'm not an expert on fluoro microscopy, and I've done very little of any type lately on account of being busier with life. "Jack Stravinsky" is more of an authority on it than I am, and he is doing some awesome work with Borrelia using Acridine Orange and a fluorescent microscope. He is "Lymedin2010" on Lymenet.org. Unfortunately, his Photobucket account needs updated to view his pictures on Lymenet, but his videos are awesome. Notice our discussion and the details about a fluorescent microscope hack using color-specific LED lights.

flash.lymenet.org/ubb/ultimatebb.php/topic/1/120458/19



Fluorescent microscopy is used for inspecting various specimens dyed with stains that contain specific fluorochromes. It's the fluorochromes in the stain that cause different parts of the specimen to glow when illuminated under the correct wavelength of light. The correct wavelength (color of light) used is different depending on the type of (fluoro) stain one is using with the specimen, and, depending on what item in the specimen one is trying to illuminate.

Fluoro microscopy can be very helpful for those of us looking at our own blood because certain fluoro stains can illuminate the infectious organisms. For our purposes, acridine orange is the easiest one to use and obtain, and is probably the cheapest. Acridine Orange (AO) stains DNA and RNA, thus making it very easy to visualize bacteria and protozoans even at 400x. Since red blood cells only stain dull green and bacteria and protozoans stain bright orange, it is very easy to pick out microorganisms, especially if one sees the correct morphology (proper shape, size, and location).

The main reason I wanted to do my own fluorescent microscopy is because I do my own Giemsa smears and it can be nearly impossible sometimes to differentiate a Babesia parasite, or an Anaplasma morulae, from a platelet. Sometimes these pathogens take on variations of morphology that make it very difficult to say with certainty what I am seeing....especially since platelets stain the same color in Giemsa as do pathogens, and platelets typically are the same size as pyriforms and morulas. But, in Acridine Orange, platelets stain a pale, whitish, light green color. So, it's very easy to tell the difference. FWIW, spirochetes (IN ANY FORM) stain green or red.

VIEW IMAGE

Here are some examples of my own work. Notice the color of the red blood cells, the platelets, the white blood cell nuclei (which contain DNA/RNA), and the pathogens:

A Neutrophil chasing a rod bacteria between red blood cells:
VIEW IMAGE

A rod bacteria on or in a red blood cell. Notice the pale, whitish, light green platelets:
VIEW IMAGE

Typical Bartonella (orange dot) on the peripheral of a red blood cell:
VIEW IMAGE

Probable Babesia parasite in a red blood cell:
VIEW IMAGE

That last picture is very likely showing a (pyriform) Babesia parasite. Notice in this next picture how much the Babesia (ringform) resembles the platelets to the right of it. This illustrates how much easier it is to discern between platelet and Babesia in a fluorescent stain versus a Giemsa stain.

VIEW IMAGE


For those interested, to get the Acridine Orange (stain) fluorochromes to cause DNA and RNA to glow red and green, the color wavelength of blue must be used to illuminate the specimen (anywhere from 435nm to 500nm is considered in the blue spectrum). In a typical vertical fluorescent microscope, you must have the correct excitation "cube" (longpass filter) to properly use Acridine Orange stain. Incidentally, most fluorescent scopes are equipped with that one. It's always best to check before buying one though!

An illustration of the wavelength spectrum of the color blue (needed for using Acridine Orange):
VIEW IMAGE

Notice the excitation wavelength specs for Acridine Orange for the illumination of DNA and RNA (in Table 2 on this page):

/www.microscopyu.com/techniques/fluorescence/nikon-fluorescence-filter-sets/blue-excitation-filter-sets

An expensive vertical fluorescent microscope is not necessary to do fluorescent microscopy, though. As long as you have a regular brightfield microscope and some Acridine Orange, you can do fluoro microscopy (with a little improvising). A blue LED light such as this...............:

/www.amazon.com/dp/B010NQMSKA/ref=sxts_bia_sr_1?pf_rd_m=ATVPDKIKX0DER&pf_rd_p=3182441022&pd_rd_wg=mtia4&pf_rd_r=2A229T6NPV74GB11VTR7&pf_rd_s=desktop-sx-top-slot&pf_rd_t=301&pd_rd_i=B010NQMSKA&pd_rd_w=CWTm3&pf_rd_i=rgb+led+bulb+60w&pd_rd_r=8F4VT17YHR0YZHTEGGS1&ie=UTF8&qid=1505418532&sr=1

or this (the higher the wattage the better!):

/www.amazon.com/Ustellar-Changing-Waterproof-Dimmable-Security/dp/B06XJ5QH9T/ref=sr_1_9?ie=UTF8&qid=1505418532&sr=8-9&keywords=rgb+led+bulb+60w

.........pointed down on the specimen should excite the fluorochromes in the Acridine Orange and light up the typical stuff. Of course, you would only use the blue LEDs for the Acridine Orange.

Here are some examples of various degrees of hacks:

www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.org.uk/mag/artjun09/ld-fluoro.html

wbg.wormbook.org/2009/12/01/using-leds-as-a-low-cost-source-to-detect-gfp-and-dsred-2/

/asymptoticdesign.wordpress.com/2012/03/10/diy-scope/

journals.plos.org/plosone/article?id=10.1371/journal.pone.0011890

Post Edited (TOOTY) : 9/14/2017 2:17:46 PM (GMT-6)


Healrom
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Date Joined Nov 2016
Total Posts : 133
   Posted 9/14/2017 4:03 PM (GMT -6)   
Tooty, if you have a darkfield condenser, you can look the thing that you think may be a platelet. When it's stained with Giemsa, it's very distinctive with DF. So you will know for sure if it's a platelet.
It's a personnal discovery ;)

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 224
   Posted 9/14/2017 5:32 PM (GMT -6)   
Healrom said...
Tooty, if you have a darkfield condenser, you can look the thing that you think may be a platelet. When it's stained with Giemsa, it's very distinctive with DF. So you will know for sure if it's a platelet.
It's a personnal discovery ;)



Interesting, Healrom!


A person could use the RGB LED lights I linked to and simply direct them at the slide on your microscope stage to do fluorescent microscopy without the vertical illuminator. This picture here illustrates it using a stereo scope, but simply use your regular compound microscope and point one of those RGB LED lights or a specific wavelength LED flashlight at the specimen from the side as shown in this pic:

VIEW IMAGE

So, some Acridine Orange and a little improvision, and WALLAH!!!....a FLUORESCENT MICROSCOPE HACK!

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 224
   Posted 9/14/2017 5:53 PM (GMT -6)   
I finally found it! I knew I had posted a link on another thread about a cheap LED fluoro hack!

Here on this thread near the bottom of this page:

www.healingwell.com/community/default.aspx?f=30&m=3834079

Pics on this thread at this link:

www.photomacrography.net/forum/viewtopic.php?t=33123&postdays=0&postorder=asc&start=0&sid=d0c291f89ebbe9c5364fc8a1b69adccb

The pics I had in mind:

VIEW IMAGE

VIEW IMAGE


No "longpass filter" should be needed as long as your (flash)light has the proper wavelength.

Healrom
Regular Member


Date Joined Nov 2016
Total Posts : 133
   Posted 9/14/2017 6:27 PM (GMT -6)   
I have the same flashlight at home ;)

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 224
   Posted 9/14/2017 7:55 PM (GMT -6)   
I also want to mention that not only are no filters really needed if you have the correct color (LED) light, no special microscope objectives are needed. Regular brightfield objective lenses (preferrably "Plan") are sufficient. The special, high-end "fluoro" objectives that are on many fluorescent scopes only enhance the fluorescent image. They don't help produce it.

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 224
   Posted 9/16/2017 11:41 AM (GMT -6)   
Healrom said...
I have the same flashlight at home ;)



Unfortunately, if you are going to use that particular flashlight, you WILL need a longpass filter because 365nm is NOT the correct wavelength to excite the fluorochromes in Acridine Orange (this fact is why that microscopist using "your" flashlight needed a longpass filter).

All you need is a light with the wavelength of 460nm-500nm. A light with this waveband will not need a longpass filter. Those color changing LED lamps I linked to from Amazon should do the trick.

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 224
   Posted 9/18/2017 3:37 PM (GMT -6)   
TOOTY said...
All you need is a light with the wavelength of 460nm-500nm. A light with this waveband will not need a longpass filter.


Theoretically this should work. I just tried it myself with a AO-stained smear but for some reason I cannot get it to work by illuminating my specimen from the top of the stage with the blue LED. I am only using the smaller light I linked to ( /www.amazon.com/dp/B010NQMSKA/ref=sxts_bia_sr_1?pf_rd_m=ATVPDKIKX0DER&pf_rd_p=3182441022&pd_rd_wg=mtia4&pf_rd_r=2A229T6NPV74GB11VTR7&pf_rd_s=desktop-sx-top-slot&pf_rd_t=301&pd_rd_i=B010NQMSKA&pd_rd_w=CWTm3&pf_rd_i=rgb+led+bulb+60w&pd_rd_r=8F4VT17YHR0YZHTEGGS1&ie=UTF8&qid=1505418532&sr=1 ), so I wonder if I don't have enough wattage. But, I can't discern even a hint of glow.

So, I'm not sure what is going on, because, like I said, theoretically it should work.

I can make it work by shining the blue LED through the back of my vertical illuminator, but only if I slide the longpass (dichroic mirror/filter) into place. Since using the dichroic mirror is the only way I can direct the light down through the objectives to the specimen, I have no way of knowing if this would work if I had a "clear" dichroic mirror to direct my blue LED light down through the objectives, and therefore down to the specimen.

I know Lymedin2010 on Lymenet.org has done a hack with LED lights, but he shone the light through the back of his vertical illuminator with the dichroic (or longpass mirror/filter) in place as I just did. So, maybe a person does need to have a vertical illuminator assembly (with dichroic mirrors), thereby making the mercury bulb and lamp housing the only dispensable items in this LED hack. So, basically what I'm saying is that a vertical fluorescent scope may be necessary to do fluorescent microscopy (just not the mercury bulbs), or at least a vertical illuminator assembly/light train that would fit your own scope.

Sorry if this post was confusing, but,

I just wanted to let you guys know before you bought any lights, and then blamed me for it not working with your brightfield scopes. smile

JohnB2
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Date Joined Sep 2017
Total Posts : 2
   Posted Yesterday 10:51 AM (GMT -6)   
Hello,

I am new to this forum and my wife has had lyme for many years. I have five kids with congenital lyme. I spent the first year studying and watching everything I could and then started microscopy over a year ago. I have been in contact with many scientists, and perhaps I am sort of an activist, but some do not like me as I have called them to task for some of their dishonesty, or unwillingness to acknowledge that borrelia constantly leaks into the bloodstream as Drs. Mysterud and Laane have taught us.

Some lyme scientists have helped me greatly and one scientist performed multiple positive borrelia DNA tests post-treatment for my wife and six-year old son. Dr. MacDonald was kind enough to give me some pointers getting started before he retired. I have also been fortunate to have one of the top L-form scientists help me for a while. As I state in my videos, she said she sees an abundance of L-forms proliferating and exhibiting all possible morphological variations. Some scientists believe that they have a concurrent B. Miyamotoi or B. Mayonii infection, or both, along with Bb, as they have many specific Bb antibodies. The truth is, they can have tens of thousands in their blood in one drop during a febrile period. Because of this, the forms generate the most morphologies. Regardless of the type, it is all borrelia. Dr. MacDonald advocated the disease being called borreliosis, not lyme. All forms of borrelia can go everywhere.

I started doing fluorescent microscopy about a number of months ago with acridine orange. I will submit a video soon that has the most spirochetes I have ever seen in any video. They are clearly pictured forming everything in this mass of thousands of borrelia. A while ago a scientist tested them for nematodes, and it was negative, and I came to realize that the spirochetal or nematode shaped forms in many peoples' blood is simply aggregates of spirochetes. Videos on my site demonstrate this, and the video I will post soon presents the most vivid evidence.

Most people do not watch more than a short part of my videos, but if you take the time to watch some, you will see many spirochets and L-bodies, which are simply spirochetal, cystic, and granular forms making various shapes. The new video I am going to post has dozens of L-body walls being formed around various numbers of spirochetes. There is so much conjecture about spirochetal morphology, but I am convinced with acridine orange and high levels of borrelia in the blood, many of you will be shocked at what it reveals. Here is my main Youtube site: Thanks!

https://www.youtube.com/channel/UCMcCeO0tG4R4HOy8fFYIEnA

Yellow Cat
Regular Member


Date Joined Dec 2016
Total Posts : 188
   Posted Yesterday 11:59 AM (GMT -6)   
@JohnB2, welcome to forum! I've been watching your videos for some time now, do you do any lyme specific treatment? I wonder how the situation will change after using antimicrobial herbs.

JohnB2
New Member


Date Joined Sep 2017
Total Posts : 2
   Posted Yesterday 11:06 PM (GMT -6)   
Hello,

We have tried various treatments, but respecting the wishes of my wife, she prefers I not discuss particulars. For my part, I prefer to stay focused on understanding the morphology in figuring out what we are up against and hoprefully someone will learn how to adequately kill L-bodies. The new video that I will post illustrates what I believe to be the most difficult aspect of borrelia infections. The L-form scientist who has been working in the field for 30+ years stated: "It is very unfortunate that inhibiting L-forms with pharmaceuticals is very difficult". There is some experimentation with some very toxic antibiotic cocktails to attempt to kill L-forms in vitro; but the success has been limited and I do not believe that success in vivo will be achieved yet. There is much talk about biofilms, and Dr. MacDonald's FISH verification of Bb being the cause of 100 Alzheimer's lesions is illustrative of the problem. Dr. MacDonald also taught about the round body brain diseases, and had many micrographs of cystic invasion of neurons and other areas of the brain. I believe that there are far more L-bodies than biofilms with borrelia, and they are incredibly hard to kill and very pathogenic. The new material I have clearly demonstrated that the many morphological shapes one sees in my videos that are the more formed, and often green or yellow-fluorescing L-bodies are all formed by spirochetes. If 80 spirochetes form an L-body, there is a problem, as the L-body itself can transfect human cells and cause many other issues, or the various morphologies within the L-body can exit.

I am not opposed to attempting many types of treatment, but I do believe that if more people were to use fluorescence with acridine orange - they would be surprised at how much borrelia they have. All of the intracellular borrelia in the RBCs will be revealed. And take for instance the Lida Mattman "long form" - acridine orange reveals in the case of my family that they are always there - in every sample. These forms are made up of many spirochetes. If every L-body is made of spirochetes or morphologies that put a cell wall around a bunch of granules(which are also spirochetes) - imagine how many spirochetes everyone really has. Most people cannot see them with the methods they are using. I couldn't see much when I first started with a regular brightfield microscope at first. Also, when people leave a blood sample on a slide for a while, they will see mostly L-forms and not spirochetes, that is why for acridine orange it is important to immediately use the AO working solution on the sample before they all morph to L-forms.

Another aspect to consider is the autopsy of Dr. MacDonald on the MN man who had been on ABX for years and anti-microbials. I understand that he lived in the woods and likely had many tick bites, but the borrelia was everywhere. Then there are all the Youtube videos like the woman from France who had been on Doxy, Metronidazle, and something else for a long time and they were all over her sample. I truly do not mean to be a pessimist, but I need to be a realist - I have six family members with this and we just had another baby who surely must have this. Drs. Mysteruud and Laane stated that the source is in the body and they constantly leak into the bloodstream. The new video demonstrates an incredible proliferation beyond belief that I will soon post. I also think that additional relapsing fever strains are more prevalent than some think. Didn't Dr. MacDonald verify more cases of B. Miyamotoi than the entire CDC as I recall? I am still looking for a reliable relapsing fever test, but unfortunately Dr. MacD's probes are no longer available.

Anyways, it is encouraging that there is a global network of scientists, activists, and those who just want to face the reality and try to do something about it.
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