For those of you doing your own microscopy... *Part 4*

New Topic Post Reply Printable Version
41 posts in this thread.
Viewing Page :
 1  2 
[ << Previous Thread | Next Thread >> ]

Mustard Seed
Veteran Member


Date Joined May 2016
Total Posts : 1168
   Posted 9/27/2017 9:26 PM (GMT -6)   
Part 1: www.healingwell.com/community/default.aspx?f=30&m=3834079
Part 2: www.healingwell.com/community/default.aspx?f=30&m=3849152
Part 3: www.healingwell.com/community/default.aspx?f=30&m=3859907

To recap the conversation:

JohnB2 said...
Hello,

I use a E200 Nikon-grade microscope with a fluorescent attachment and acridine orange. I could barely see anything with a cheap microscope initially in brightfield. Most videos I have seen online that people do will never see all these forms. Regular brightfield will not work well and many spirochetes are inside the RBCs. Phase contrast provided incredible ability to learn about how these morph, which I did for about a year. I would go back and read the science info after I would notice various elements in phase contrast. This form is very common and you can find it all over online, not just in scientific journals, but even an generic Google images searches.

Acridine orange will kill these bacteria. They are dead, and they fluoresce with RNA especially when they are in their aggregation and building mode. When the L-bodies made up of spirochetes become more cohesive, the typically fluoresce green or yellow expressing DNA. It doesn't matter that they are dead, as the L-form scientist who I corresponded with told me as long as I could get a good micrograph. When you have caught many of the forms that Dr. Mattman and MacDonald described, and when you start to have way more known morphologies than you can find in scientific journals, than I can see what the L-form scientist meant. Borrelia L-forms are unique among L-forms, as my videos point out by quoting Dr. Mattman. Unlike other L-forms, they often have spirochetal forms protruding rather than infinite varieties of rhizoid forms.

Acridine orange has opened up a whole new world to me. I have never seen anyone with multi-spirochetal cysts, other than in scientific journals before I found 100's of them. This is not because of anything I did, but I believe that many of you would find exactly the same things. We are entering taboo-land now. Why if this is so easy and acridine orange has been around for decades, do we not have a scientific catologue with tens of thousands of images? They don't want us to realize that these are so easy to find. Yes, I grant that my family may have B. Miyamoti, B. Mayonii, additionally, but all borrelia is by definition relapsing fever bacteria due to antigenic variation. However, with some blood drops of my family with tens of thousands of microbes, I think multiple kinds of borrelia is probable.

I never stopped and asked - what makes up an L-form when I first started doing this. But obviously with the case of borrelia, spirochetes build them. The spirochetes themselves aggregate, inflate and form the L-body, or spirochetes work together to put a thin wall around other members of the aggregate.

I have made some videos about the Mattman long form L-body, that many mistake for a nematode. I thought they were nematodes too, but I had a scientist do DNA testing for nematodes and it was negative. Then many nematode scientists watched some video, and some admitted they look like nematodes, but they said that they are definitely not. I kept studying them with acridine orange, and many looked just like the smaller L-bodies, with spirochetal morphologies inside and protruding from them. This is exactly how Dr. Mattman described the borrelia L-body. Finally, I caught the preliminary stages of borrelia "Mattman long L-form" building and they were made up of spirochetes building a large L-body that was spirochetal shape. They do this all the time. I thought to myself - "all these years there is nothing about these from scientists" since Dr. Mattman pointed them out? They are the easiest diagnostic aid imaginable - how could they miss this target with borrelia antibodies or as an indicator for preliminary diagnostic screening that results in further testing? These Mattman long forms are taboo-land again. But they are in every single sample I test - many of them. Borrelia is arrogant and it likes to form a large shape like itself. I am going to post a short video with a couple of these Mattman long forms that were in the single drop of blood that you just watched a video of - one is loaded with spirochetal morphologies and one is building a cohesive long L-form, where part of it is spirochetes aggregating to build the corpus, and part of it is fluorescing green.

I do hope that many others will start to use acridine orange.

Here's a link to JohnB2's YouTube: /www.youtube.com/channel/UCMcCeO0tG4R4HOy8fFYIEnA

Jackss said...
Yes, that is a spirochete that is drilling very fast in the plasma & around red blood cells of someone who is chronically ill for 40 years. Even in ticks some spiros can drill this fast.

John B, Acridine Orange can also stain indiscriminately & I had seen your AO stain in the past & those are not spirochetes. What happens is that the wbc's lyse & get strewn all over in the smear & you will get string-like subjects that are really part of the DNA from the Wbc's.For you to get a better idea, stain a normal persons blood & you will see the same artifacts. Full strength AO will be cytotoxic to Borrelia & can destroy most of them. Lab workers have also reported that full strength AO will also lyse sperm when they tried it & so they know it is toxic to certain cells in it's purer forms. Spirochetes are hardy yet delicate creatures.

I use live cell AO staining, in which I dilute AO .2% with NaCl (Salt water) & I give the exact dilution method that I used in the description of my video. Too concentrated & it can kill the spiros, not concentrated enough & one can miss the spiros if they are there. Sometimes even when you get the concentration right even the amount of AO drop that you use with the blood drop can alter the outcome, as it is hard to control the exact amount of a drop without a pipette.

When you look for AO stains in live preps, look for the known shapes, movements & morphologies (SOP or Tennis Rackets...etc) & not just any strings or string conglomerates. I hope this helps, as I want you to be successful too...all the best and if you need any help just LMK.

String of Pearls & 2 daughter splitting spiros in this video can be seen, as well as blebs/granular forms. Not all the green dots are spiros though, as some are from wbc components (lysosomes & peroxisomes).
https://www.youtube.com/watch?v=qlL8ZLitiBI

Continue:

Girlie
Forum Moderator


Date Joined May 2014
Total Posts : 32624
   Posted 9/27/2017 9:29 PM (GMT -6)   
Thanks MS - you read my mind.
Moderator, Lyme Forum
Symp started April/2013; Buhner's Lyme May 15-July24/14; Igenex pos. July 3/14
Doxy: July 4-Aug.24/14;Zithro July26-Aug24/14; Amox + Proben. Aug. 29/14;
added biaxin Sept. 26/14
Disc. amox,added Ceftin Nov. 20th.;
Disc. biaxin added Buhner bart herbs Dec/14;Jan/15 pulsing Tinda (w/ Ceftin);
Abx/herb break Apr-July/15; July-mino; Aug. added Rif;
Nov./15 mino - to biaxi

Pirouette
Veteran Member


Date Joined Mar 2014
Total Posts : 6201
   Posted 9/28/2017 10:18 AM (GMT -6)   
I haven't been following this discussion closely but just want to thank you all for it - my hope is that I can come back to it when I have more time...

-p
Lyme Moderator
Chronic late-stage lyme—likely infected in '00; Clinically dx Mar'14 w/ Babs, Fry Labs+ Bart-like, CDC+ Bb. First treated 4-5 viruses & GI/immune. Herbal antimicrobials in May; IV port-started Rocephin in Nov; added vancomycin Mar'16;
DETOX: Pinella/Burbur/Parsley/Milk thistle/Burdock/Red root; Samento/Banderol/Enula; JK/Turmeric; BFM-1; antifung; many many supps; cholestyramine!

Lapis_29
Veteran Member


Date Joined Sep 2017
Total Posts : 844
   Posted 9/29/2017 12:14 PM (GMT -6)   
would still really love to see an experiment showing how a spiro might react to a low level of DMSO.

also, anyone got a rife machine? would love to see how a spiro might react to specific rife freqs. that would be fascinating!

Mustard Seed
Veteran Member


Date Joined May 2016
Total Posts : 1168
   Posted 9/29/2017 3:55 PM (GMT -6)   
How Borrelia reacts to a Rife machine is something I've always wanted to see. I'm a skeptic at heart so I would love to see some Borrelia dying in real time under magnification.

I just started doing darkfield today. I got an oil darkfield condenser and a 100x infinity lens with iris. It's sooo cool. I highly recommend it to anyone who hasn't tried it.

I'll hopefully post some pictures by the end of the weekend.

Mustard Seed
Veteran Member


Date Joined May 2016
Total Posts : 1168
   Posted 10/28/2017 10:08 AM (GMT -6)   
Found a TON of spirochetes in my blood yesterday, no incubation time needed. Here's one:

/gfycat.com/HomelyHarmoniousAddax

Girlie
Forum Moderator


Date Joined May 2014
Total Posts : 32624
   Posted 10/28/2017 12:25 PM (GMT -6)   
MS - first - you have treated with abx for awhile, right? And there's all these spirochetes floating around. (I'm assuming those squiggly 'threads' are the spirochetes)

Okay - this is going to sound 'dumb' - but are those circular 'things' the red blood cells? And the spirochetes are floating outside of the blood cells?
Moderator, Lyme Forum
Symp started April/2013; Buhner's Lyme May 15-July24/14; Igenex pos. July 3/14
Doxy: July 4-Aug.24/14;Zithro July26-Aug24/14; Amox + Proben. Aug. 29/14;
added biaxin Sept. 26/14
Disc. amox,added Ceftin Nov. 20th.;
Disc. biaxin added Buhner bart herbs Dec/14;Jan/15 pulsing Tinda (w/ Ceftin);
Abx/herb break Apr-July/15; July-mino; Aug. added Rif;
Nov./15 mino - to biaxi

Mustard Seed
Veteran Member


Date Joined May 2016
Total Posts : 1168
   Posted 10/28/2017 12:46 PM (GMT -6)   
Yes all correct. The blood cells are usually more rounded but they've be spread under a cover slip here so they become a spiked and crenated.

Girlie
Forum Moderator


Date Joined May 2014
Total Posts : 32624
   Posted 10/28/2017 12:59 PM (GMT -6)   
So what do you make of all the lyme still in your blood?

What does your LL Dr. say?


That's discouraging to say the least!!!
Moderator, Lyme Forum
Symp started April/2013; Buhner's Lyme May 15-July24/14; Igenex pos. July 3/14
Doxy: July 4-Aug.24/14;Zithro July26-Aug24/14; Amox + Proben. Aug. 29/14;
added biaxin Sept. 26/14
Disc. amox,added Ceftin Nov. 20th.;
Disc. biaxin added Buhner bart herbs Dec/14;Jan/15 pulsing Tinda (w/ Ceftin);
Abx/herb break Apr-July/15; July-mino; Aug. added Rif;
Nov./15 mino - to biaxi

gfields
Veteran Member


Date Joined Oct 2015
Total Posts : 956
   Posted 10/28/2017 1:28 PM (GMT -6)   
What kind of microscopes and cameras are you guys using?

Lapis_29
Veteran Member


Date Joined Sep 2017
Total Posts : 844
   Posted 10/28/2017 2:04 PM (GMT -6)   
Mustard Seed said...
Yes all correct. The blood cells are usually more rounded but they've be spread under a cover slip here so they become a spiked and crenated.


suggestion

take a blood sample, view it. then drink a gatorade with 500 mg potassium dissovled in it. wait, then take another sample. you will be amazed at the difference.

Mustard Seed
Veteran Member


Date Joined May 2016
Total Posts : 1168
   Posted 10/28/2017 3:43 PM (GMT -6)   
Girlie said...
So what do you make of all the lyme still in your blood?

What does your LL Dr. say?


That's discouraging to say the least!!!

Not too much to make of it, the buggers are hardy; though I'm not treating right now. My LLMD hasn't seen any microscope videos, but we've talked about it and she found it very interesting. We talked about how some people pay to get darkfield microscopy done on their blood. I spent too much time treating co-infections I think. I haven't found any evidence of Babesia or Bartonella in my blood. The strongest antibiotic regime I did for Lyme was probably 3 months of Rifampin/Biaxin/Minocycline followed by 3 months of Rifampin/Biaxin/Tindamax, all while doing Buhner.

gfields said...
What kind of microscopes and cameras are you guys using?

Mine is an American Optical 10 Series. I'm using the original illuminator (18 or 20w I believe), but it was never meant for darkfield at 100x (under-powered), which is why my video's contrast is so poor. I've done darkfield with a 63x objective which yields a better image, but in my opinion, the drop in magnification isn't worth the gain in contrast. The 100x objective is an AO Spencer 100x 1.25NA with adjustable iris. The camera is an iPhone 5c with a mount for the ocular I purchased on Amazon.

Lapis_29 said...
suggestion

take a blood sample, view it. then drink a gatorade with 500 mg potassium dissovled in it. wait, then take another sample. you will be amazed at the difference.

Thanks I'll try that sometime. The weird looking red blood cells don't really bug me too much.

Girlie
Forum Moderator


Date Joined May 2014
Total Posts : 32624
   Posted 10/28/2017 4:31 PM (GMT -6)   
Mustard Seed said...
Girlie said...
So what do you make of all the lyme still in your blood?

What does your LL Dr. say?


That's discouraging to say the least!!!

Not too much to make of it, the buggers are hardy; though I'm not treating right now. My LLMD hasn't seen any microscope videos, but we've talked about it and she found it very interesting. We talked about how some people pay to get darkfield microscopy done on their blood. I spent too much time treating co-infections I think. I haven't found any evidence of Babesia or Bartonella in my blood. The strongest antibiotic regime I did for Lyme was probably 3 months of Rifampin/Biaxin/Minocycline followed by 3 months of Rifampin/Biaxin/Tindamax, all while doing Buhner.




So 6 months hitting it hard...and still there! Good thing the Bart (and Babesia) protocols also target lyme.
Jeesh!

I'm thinking of doing another PCR to test for Lyme, Bart and Babs again. $125 per infection - includes urine and blood.
Moderator, Lyme Forum
Symp started April/2013; Buhner's Lyme May 15-July24/14; Igenex pos. July 3/14
Doxy: July 4-Aug.24/14;Zithro July26-Aug24/14; Amox + Proben. Aug. 29/14;
added biaxin Sept. 26/14
Disc. amox,added Ceftin Nov. 20th.;
Disc. biaxin added Buhner bart herbs Dec/14;Jan/15 pulsing Tinda (w/ Ceftin);
Abx/herb break Apr-July/15; July-mino; Aug. added Rif;
Nov./15 mino - to biaxi

bluelyme
Veteran Member


Date Joined Nov 2015
Total Posts : 4715
   Posted 10/31/2017 11:51 PM (GMT -6)   
gfields said...
What kind of microscopes and cameras are you guys using?


im using a old university wesco and a ancient bauch and lomb .both cl scores ...i have phone holder but the cheap amazon cam with tube extension to get vga out to computer screen, then video that but its grainy.

got to do a test run of amscope dk490 . it was ok , not clean on arrival but to do darkfield at 1000x one would need a different condenser and a oil lens $...
400 x with 2x tube or 20x eyepieces(800x) was ok

Lapis_29
Veteran Member


Date Joined Sep 2017
Total Posts : 844
   Posted 11/2/2017 7:14 PM (GMT -6)   
can someone tell me if this scope will work to view spirochetes?

thank you

OMAX 40X-2500X Lab Binocular Compound LED Microscope with Double Layer Mechanical Stage and Digital Camera

/www.amazon.com/OMAX-40X-2500X-Binocular-Microscope-Mechanical/dp/B00SGCDYYI/ref=sr_1_4?ie=UTF8&qid=1509671863&sr=8-4&keywords=darkfield+microscope+with+camera&dpid=51MagaNax1L&preST=_SX342_QL70_&dpSrc=srch

bump

Post Edited (Lapis_29) : 11/4/2017 8:10:28 PM (GMT-6)


Lapis_29
Veteran Member


Date Joined Sep 2017
Total Posts : 844
   Posted 11/6/2017 12:33 PM (GMT -6)   
bump

any feedback on my microscope ? above?

Mustard Seed
Veteran Member


Date Joined May 2016
Total Posts : 1168
   Posted 11/6/2017 1:40 PM (GMT -6)   
It doesn't say whether the condenser is an oil condenser or dry condenser. If it's dry, you'll be limited to the 40x objective, and you won't see spirochetes with that in brightfield. If it's an oil condenser, you can use the 100x objective, and might be able to spot some spirochetes with practice, but no where near as good as darkfield.

My recommendation is if you're looking for spirochetes, aim for darkfield with a 100x, and if you're looking for coinfections, use 100x in brightfield with a quick dip stain.

Healrom
Regular Member


Date Joined Nov 2016
Total Posts : 158
   Posted 12/20/2017 1:15 PM (GMT -6)   
Congrats for your microscope mustard seed yeah

You have a very nice quality! The videos made with good quality smartphones renders well.

It is not surprising to see spirochetes after a treatment. They can come back the NEXT DAY when you stop it, if you are loaded.
Even worse than that, you can even see spirochetes during a strong antibiotics treatment... It was my case...

The only things that get rid of it for me are the buhner protocol (cat's claw and andrographis mainly), ozonated water, and maybe amoxicillin at high doses (but I haven't tested all antibiotics, like bactrim or rocephine)

Psilociraptor
Veteran Member


Date Joined Jul 2016
Total Posts : 1228
   Posted 12/20/2017 5:42 PM (GMT -6)   
Mustard Seed said...
.

My recommendation is if you're looking for spirochetes, aim for darkfield with a 100x, and if you're looking for coinfections, use 100x in brightfield with a quick dip stain.


This. I personally love my Omax even though some people like to poopoo the brand. But one thing I do regret is not getting a 100x darkfield. I went with the combo which had brightfield, phase contrast, and darkfield. And while cool, it doesn't have 100x on phase contrast or darkfield which seriously limits your ability to see these bacteria. I would have rather gone with one or the other all the way. You can still see spirochetes in a good sample, just way harder to make out

Healrom
Regular Member


Date Joined Nov 2016
Total Posts : 158
   Posted 12/21/2017 10:18 AM (GMT -6)   
You can still buy a second hand one. I've had mine for 50 euros.
It's a 100x glycerin immersion.

What you have to take care is not taking an infinite objective, but a 160. (if your microscope is 160, but I guess it is)

bluelyme
Veteran Member


Date Joined Nov 2015
Total Posts : 4715
   Posted 1/22/2018 11:58 PM (GMT -6)   
/youtu.be/pS1QjhVAEa0

Bumping with our french friends scope work of some ketes in a urine sample

And with news that blue got a decent phase contrast ! On extended loaner ..superexcited pocketprotector here

bluelyme
Veteran Member


Date Joined Nov 2015
Total Posts : 4715
   Posted 2/13/2018 4:24 AM (GMT -6)   
Keep up the good work johnb2 . They cant front on the truth !

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 262
   Posted 2/13/2018 6:11 AM (GMT -6)   
I don't check the forums as much anymore but the microscopy threads always catch my eye when they pop up. I'm glad to see more patients getting involved with microscopy. I personally have not had as much time lately. I'm really glad to see you got a phase scope Bluelyme!!!

I'm just popping in here to mention that debris and artifacts can indeed stain in acridine orange. They can be mistaken for atypical borrelia morphologies. So, watch out for that.

Another thing to mention is that acridine orange is clinically used to evaluate red blood cell morphology, such as nuclear remnants. Nuclear remnants can appear as nondescript red cell pathologies when in fact there is no infection. The main advantage with acridine orange is to view known morphological pathologies. I would be hesitant to label objects in dried smears that don't have distinct comparable morphologies unless one is using fluorescent DNA-specific stains. The same for wet mounts unless distinct morphological transformation has taken place (under the scope) or known morphology is seen such as distinct, typical l-form borrelia strings or their gemma cysts.

Just because something glows with acridine orange does not mean it is pathogen-related. ONLY with DNA-specific stains such as antibody stain is it always true that what glows is pathogen-related. That is what Dr. Alan MacDonald was working with, not acridine orange.

TOOTY
Regular Member


Date Joined Apr 2014
Total Posts : 262
   Posted 2/19/2018 9:57 PM (GMT -6)   
I am not here to try to discredit anyone, John. I agree with you that microscopy gives us more than just a guess at what we are dealing with! I would just like more verification in your videos about what you are proposing you are seeing.

Like I said, acridine orange indiscriminately stains all DNA/RNA, and unfortunately even stains some non-living things like contaminates. There is a HUGE difference between acridine orange stain and the specific fluorescent stains that Alan MacDonald and these other scientists were using!!! HUGE difference. It is not fair to compare micrographs of the two unless there is CLEAR MORPHOLOGY and MAGNIFICATION COMPARISONS.

I am definitely not suggesting that anything you show is contaminates, but the unfortunate fact for many getting started (not you) with blood microscopy is that at times what they are seeking info on is in fact debris such as dust, fibers, or even microscopic hairs. Particularly the samples people think may be filarial worms. I've seen that time and time again.

Your repertoire of microscopic illustrations from some of the leading Borrelia scientists/pathologists is quite amazing. The trouble I personally have with your videos is that many of your pictures are flashed so fast from frame to frame it's hard to see the context. Especially since there are no magnification comparisons. I have nothing wherewith to judge the magnification of YOUR material compared to the micrographs you show from the scientists. And, since Acridine Orange indiscriminately stains all DNA/RNA and some non-living things, I have no way of knowing if what you are showing is what the scientists were showing.

Eaglet
Regular Member


Date Joined Nov 2017
Total Posts : 44
   Posted 2/20/2018 4:53 PM (GMT -6)   
Any tips on how could I share my microscopy pictures/videos here on Healingwell? Where could I post them first?

There were spirochete-like things moving like spirochetes tend to do... I just would like some confirmation.
New Topic Post Reply Printable Version
41 posts in this thread.
Viewing Page :
 1  2 
Forum Information
Currently it is Friday, June 22, 2018 1:30 PM (GMT -6)
There are a total of 2,974,538 posts in 326,186 threads.
View Active Threads


Who's Online
This forum has 161253 registered members. Please welcome our newest member, A_Warrior's Queen.
385 Guest(s), 11 Registered Member(s) are currently online.  Details
Foo223, mattamx, lymenc4, Girlie, Pratoman, Hibee, hrpufnstuf, DJBearGuy, LJohn23, RunJerRun, U.C.Me?