Lymedin2010
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posted
Glad to see more are interested.
Scopes & what works for one person can be very subjective, as some choose to spend the minimal amount & are happy to just see spirochetes. Others will not be content to simply see them, as they will want sharper & clearer images & video, and yet others will require upper echelon scopes that are more sensitive to pickup fluorescent stains.
Most lab grade scope will be just fine to view blood & spiros.
"Plan" objective lenses means the whole field is in focus & we like these lenses since we don't desire blurry edges & vignetting.
Generally achromatic lenses are good enough & I use those (plan achro). But apochromatic are better & are optimized for 3 colors coming together simultaneously, as opposed to only 1 color (2 elements: an achromatic lens and a meniscus lens) for the achromatic & 2 colors for the fluorites objectives. The plan-apochromatic are the most complex & can have up to 15 elements & are far more expensive. Normally better to worst type objectives (apochromatic -> semi- apochromatic -> achromatic -> non-achromatic), which also means pricier -> cheaper. . These can also dependent upon the application needed. Achromatic is the norm & what usually microscopes are fitted with, as they provide a relatively good price to acceptability point. The differences between apochromatic & achromatic objectives quality can sometimes appear subtle & all depending on the preparation of the objects being viewed & application (ex. microphotography or fluorescent capture).
For the condensor, typically the higher the NA the better the image, but one can get by with a NA of 0.9 & get good results. My Zeiss is a NA 0.9 & the Reichert even worst as it is an Abbe & not even achromatic, but can pickup far more detail due to oblique illumination & so sometimes it is not always about the numbers or types. The trade-off is because of the Abbe the images are not as sharp as they would be with achromatic or apochromatic.
Lymedin2010
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Ask about the objectives on the Reichert. My Reichert 40x is comparable to my Zeiss 40x, but my NPL Fluotar on the Zeiss blows the Reichert Plan Achro out of the water & is far more expensive as well. You can still see the spiros with 100x oil on the Reichert, but they will not look as crisp & sharp as the Zeiss & some people will be ok with this.
My Reichert: Plan Achro 40x (Plan Achro 40/0.66 ph, infinity/0.17 1734) Plan Achro 100x (Reichert USA 100, 1736)
Keep in mind that sometimes the preparation itself can also make the difference between video comparisons as well as the actual glass used.
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quote:Originally posted by Lymedin2010: Ask about the objectives on the Reichert. My Reichert 40x is comparable to my Zeiss 40x, but my NPL Fluotar on the Zeiss blows the Reichert Plan Achro out of the water & is far more expensive as well. You can still see the spiros with 100x oil on the Reichert, but they will not look as crisp & sharp as the Zeiss & some people will be ok with this.
My Reichert: Plan Achro 40x (Plan Achro 40/0.66 ph, infinity/0.17 1734) Plan Achro 100x (Reichert USA 100, 1736)
Keep in mind that sometimes the preparation itself can also make the difference between video comparisons as well as the actual glass used.
Love your videos Lymedin. Actually it was your Youtube video where you show how to do a blood smear that got the wheels turning in my head for all this. So thanks a ton, I find it so cool!
Ya that Zeiss you linked looks pretty nice. Too bad I'm a day too late.
I would like to experiment with darkfield or phase contrast, so I'm considering buying something that has those objectives/condensers ready to. So it's a trade-off between either getting a really nice lightfield scope and upgrading later if I want (more $), or finding something a little less nice but with darkfield and/or phase contrast (I've seen some AO 10's with phase).
Lots more researching and hunting around for me!
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TNT
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quote:Originally posted by mustardseed2:
Love your videos Lymedin. Actually it was your Youtube video where you show how to do a blood smear that got the wheels turning in my head for all this. So thanks a ton, I find it so cool!
Yes, Lymedin is doing AWESOME work! He has been the inspiration for many of us.
quote:Originally posted by mustardseed2: I would like to experiment with darkfield or phase contrast, so I'm considering buying something that has those objectives/condensers ready to. So it's a trade-off between either getting a really nice lightfield scope and upgrading later if I want (more $), or finding something a little less nice but with darkfield and/or phase contrast (I've seen some AO 10's with phase).
Lots more researching and hunting around for me!
That's the predicament many of us suffering from, and treating for lyme, are in. But, I think what we are all saying is that you can start with pretty much anything-but to start out with as nice a scope as one can afford- and upgrade as you are able. The pocketbook is the limiting factor when it comes to microscopes.
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Lymedin2010
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I'll check for a darkfield from time to time & post anything if I see it.
If we ever get any skin lesions or cuts, it might be worth looking at broken scab bleeds & surface lesions under our scopes.
"Background Necrobiotic xanthogranuloma (NXG) is a rare histiocytic disorder of unknown origin. Objective We conducted an investigation of skin biopsy specimens from 7 patients with NXG for the presence of Borrelia by focus-floating microscopy.
Results Borrelia could be detected as single, paired, or clusters of spirochetes in 6 cases of NXG whereas two cases investigated with a Borrelia-specific polymerase chain reaction (23s-RNA) remained negative."
This means FFM is better at detecting Borrelia than PCR!!!
Lymedin2010
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I captured another cyst formation. The transition goes from atypical spiro -> to "Tennis Racket" form -> then to cyst. Check the Youtube description for detail.
Full morphological sequence in this video: Atypical form -> "Gun Holster" -> "Tennis Racket" -> Cyst.
Now 3x Lymies with nematodes in their blood. This on top of Alan MacDonald studies on nematodes in MS patient autopsy.
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posted
I am sick with flu today, but I did see a similar" worm" in live blood a few weeks ago-- when I get up to it I will try to post the pic here if it is not too dried out.
The worm in the video above seems to be about 20 to 30 times longer than the average red blood cell--- so we can rough ball the size. It also seems to be somewhat flat and pointed at both ends. Do you think its similar to these filariaforms? Posts: 696 | From New York | Registered: Aug 2006
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quote:Originally posted by Lymedin2010: I captured another cyst formation. The transition goes from atypical spiro -> to "Tennis Racket" form -> then to cyst. Check the Youtube description for detail.
Full morphological sequence in this video: Atypical form -> "Gun Holster" -> "Tennis Racket" -> Cyst.
Amazing videos Lymedin2010--- it looks like that spiro which was hanging off the red blood cell encysted directly INTO the red blood cell near the end of the video.... hmm.. its like it catapulted itself inside while it was in the process of forming a cyst. I guess the spiro would get double protection from that maneuver-- at least for the life of the red blood cell.
Our red blood cells are like sitting ducks waiting to be parasitized and slaughtered!! If there were just a drug that could relax the spiro's sucker-like grip on the red cell wall--- that would be of major benefit to Lyme patients. Pumpkin seed oil apparently does this for worm parasites in the intestines-- they relax their grip on the intestinal wall for a few hours, and can be flushed away by eating a lot of fiber and cascara sagrada shortly after taking the pumpkin seed oil..
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Lymedin2010
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The filarial worms I have seen in ticks have a more rounded & blend end, & in our blood they appear more sharply pointed. So I do think they are another type, probably opportunistic from mosquito bites post chronic LD. Someone suggest that mine are Ascaris Lumbricoides larva, but I am not so sure.
Thanks. The time lapse runs longer before it encysts & the rbc was dragged by the moving plasma, but was held on tightly by the sticky end to the slide. I always said that the secret to killing these things is in blocking the tip entrance & in so doing we probably block the blebs too. The blebs are also generated by the tips of these spiros & so to me I think they are one the same.
The rbc's cannot simply allow things to flow intracellularly, otherwise we would not oxygenate properly & so the spiros are well protected there. IV abx would work best, as it worked for me when I first got sick & I could not see anything in my blood.
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quote:Originally posted by Lymedin2010: The filarial worms I have seen in ticks have a more rounded & blend end, & in our blood they appear more sharply pointed. So I do think they are another type, probably opportunistic from mosquito bites post chronic LD. Someone suggest that mine are Ascaris Lumbricoides larva, but I am not so sure.
Thanks. The time lapse runs longer before it encysts & the rbc was dragged by the moving plasma, but was held on tightly by the sticky end to the slide. I always said that the secret to killing these things is in blocking the tip entrance & in so doing we probably block the blebs too. The blebs are also generated by the tips of these spiros & so to me I think they are one the same.
The rbc's cannot simply allow things to flow intracellularly, otherwise we would not oxygenate properly & so the spiros are well protected there. IV abx would work best, as it worked for me when I first got sick & I could not see anything in my blood.
Lymedin2010--- this video of the worms inside a tick is particularly valuable!! We should raise some money on Gofundme and offer a $5,000 reward at college campuses in the Northeast (where ticks are easy to get --- just drag a sheet out in long grass) for definitive ID of these worms from a biology PhD student or professor. I guess we would need to do a PCR or some other genetic test to properly identify these filarial worms!! Wow-- that video shows the tick is literally stuffed with those worms. I just can't believe that the government has not bothered to ID these worms yet-- after more than 30 years.
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Eva Sapi did say that the worms had genetic elements of onchocerca, but were not onchocerca.
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This recent study from 2014 in a journal of Veterinary studies, with Eva Sapi as an author, confirms the worms to be of the Acanthocheilonema genus::
Here's a pic of the Acanthocheilonema worm lifecycle--- it has both ROUNDED and POINTED ends at different part of its lifecycle--it is rounded in the tick-- which explains the rounded worms in your video---, and it is more pointed when it is in the mouse (mammal)!!: Posts: 696 | From New York | Registered: Aug 2006
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Unfortunately for us, this worm does not harbor the symbiotic Wolbachia bacteria, which means that it cannot be killed by the antibiotics that kill wolbachia.
Lymedin2010
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Great links, thanks!
I am sure we will ultimately learn that a few different species can be transmitted in ticks. We are starting to realize that post chronic Lyme, that we can acquire further opportunistic parasitic & other forms of infections. These can be transmitted via various alternate means, such as mosquito bites, eating raw meats, swallowing a bug accidentally...etc.
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shows that an extract of the Lantana Camara plant (a pretty flower) kills 80% of the Acanthocheilonema worms and sterilizes the rest. Unfortunately i think the Lantana Camara plant is toxic to grazing livestock.
(Interesting that withania somnifera, turmeric and garlic are on the above list...) I will add these filariacidal plants to my list of Spirochetcidal compounds-- and move further links over there for future reference. Cheers...
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Lymedin2010
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Good find.
"PUMPKIN SEED (CUCURBITA PEPO) Medicinal use and Health Benefits
Pumpkin seed is used to get rid off intestinal parasites, such as pinworms and, for a long time, widely used to extract taenias or tapeworms from the human body. In the first case, it is advised to eat peeled seeds in the desired amount. In the second case, it is recommended to do the following procedure: Smash 50 gr. of fresh seeds; mix them with sugar or honey. Eat the mixture as the only food for a day in the main meals. After some hours, try to make a deposition and see if the parasite has been expelled. On the contrary, this process can be repeated on another occasion. Laxative: It favours the intestinal transit, being specially interesting the fact that it does not irritate the intestinal tract (Look at the pumpkin as an edible fruit) Anti-prostatic: Recent studies seem to suggest it is very effective, when combined with the lipophilic extract of the palm " Serenoa repens " for the treatment of benign prostate hyperplasia. By decreasing the inflammation, this gland doesn't make so much pressure on the urethra, which makes easier the expulsion of urine."
posted
Has anyone done Geimsa stains on themselves? I think all you need to do is get some blood on a slide, fix it (apparently by soaking in alcohol then drying), applying the stain, waiting, and then looking (perhaps with a wash of excess reagent).
Apparently this would find all kinds of protozoans and worms. This could save money as apparently Geimsa stains have to be done at different points over time and even different times of the day to find specific organisms.
Is this just a bad idea for someone who is not skilled in laboratory pathology?
LOL, and how crazy would your doctor think you were if you showed up with your own microscopy pictures?
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posted
How do you know a spirochete seen in your blood is not a dental spirochete (treponema genus) that got into blood from brushing teeth or flossing?
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bluelyme
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I just did a giemsa stain and saw some interesting shtuff on my friends amscope ...a few dots but one ufo that took up whole rbc..i will try to post pic ...it was quik dip vet stain thanks lyme net ...
also ohio there are 100 species of borreliosis and then there is leptospirosis, syphilis and others but even if it is dental gone systemic treatmwnt is still the same ..
Also i have my blood parasitized with toxo or proto or babs but tx is all the same .i guess for frequencys i wanna know what species but i have gotten more answers from a scope that 33 ducs
lymed did a vid of his mouth stctuff a while back..also a dentist named norquist has done good u toob vids of spirochetes using h202 ,clorone dioxide ,and others his best results were laser and honey with herbs...
personally propolis has helped gums ..too bad i cat do that iv...ha
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bluelyme
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These pics are bad but i was stoked to do the stain..i likely dont have enough light using my buddies scope ..first pic shows weird artifact anybody seen such a creature?
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TNT
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quote:Originally posted by bluelyme: These pics are bad but i was stoked to do the stain..i likely dont have enough light using my buddies scope ..first pic shows weird artifact anybody seen such a creature?
My first thought and opinion is that it resembles Babesia canis (Babesia that generally infects dogs).
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TNT
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The size and shape of the organisms in the RBCs of the first and third pics seriously resemble Babesia canis.
The size and shape of the dots on the RBCs of the second pic suggest Bartonella.
[ 06-15-2016, 09:21 PM: Message edited by: TNT ]
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bluelyme
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Thanks tnt ..yes on the bart, on the other ,the lab at african university is trying to say its toxoplasmosis.i was thinking proto but now that you mention the canis i have been bit by a few dogs..
..same sorta treatment was just hoping to narrow it down for frequencys. And yes ohio i showed a nurse at the er my spirochettes video and he was facinated but in disbelief. My tcm rife duc takes a it at face value..
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Lymedin2010
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Word on the street is that a lot of times the rbc may have inclusions within the cells that look just like little dots & a doctor had told us to be weary of this. I would prefer to see the other forms of babs for a more guaranteed positive ID.
I captured another SOP reverting back to an atypical form. I think the large SOP's may be rbc wall phospholipid formations & I will have some good proof video clips once I put it all together in a future video. https://www.youtube.com/watch?v=FKZLVwqe_fk
I also include a LIVE VIEW of the same video above, to show that the pearls are really gone & have been consumed by the body & to dispel any notions of focus issues (in case anyone questions it). https://www.youtube.com/watch?v=K5bR3D_b5xs
TNT
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quote:Originally posted by Lymedin2010: I would prefer to see the other forms of babs for a more guaranteed positive ID.
I really lean towards the B.canis. If followup giemsa smears begin to show a clearing in the pyriform shapes, I would feel 100% positive.
If his original smears were stained too long, that could possibly make them dark like that.
Blue, I would continue to monitor your blood via Giemsa stain and look for more items like what you showed us, especially a clearing in the pyriform shapes, or other Babs morphologies.
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Lymedin2010
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There was a nice article the doc posted on the inclusions, I will have to find it & some pitfalls to watch out for when staining.
***Atypical spiro to "Tennis Racket" form & then to Cyst (Discoid Gemma type) directly in HUMAN BLOOD. Tennis racket forms are widely known & seen in Borrelia obtained from ticks & wild grown in culture. https://www.youtube.com/watch?v=Eg_Id74a_4I
Lymedin2010
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Sweet deal for someone...I love those types of deals. As I said before I got 2x microscopes for $200. Just the 1000x objective on one of the scopes was really worth $300-400.
[ 06-26-2016, 04:10 PM: Message edited by: Lymedin2010 ]
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Lymedin2010
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My blood on BSK-H on day 5. I froze the blood sample to lyse most of the rbc's & hopefully release the true spiros & eliminate any false spiros.
posted
Im not sure if this has been asked before, however does anyone offer assistance reviewing members blood for tbd here? I am really interested in this type of study but not sure if I am able to complete the analysis myself or work the equipment given the severity of my neurological illness right now. Thanks!
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bluelyme
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Kms lymed posted this in another thread ...you may be able to find a practioner here ..also there is duc here who will send samples to africa for darkfield identification ..not cheap
I am in buying mode for a new scope. Looking for a model with high resolution at 1000x and greater. Fluorescence capability is a definite plus, but one that is convertible is good as well.
Maybe y'all know some good brands/models. Price is not a concern. Dont care if it's 10,000 dollars. I am going all in.
Let me know if you see any good deals on ebay, or know of models straight from manufacturer.
It's hard to find example images or.video from any models, so I will certainly use the return policy to my advantage to make sure it is a good fit.
Time to upgrade from my cheap amscope.
Once I find a model, my current setup will also be for a sale, and Id let it go for a very reasonable amount for anyone here that wants it. As in beat any price and adjust based on financial situation, just to share the capability to do this yourself.
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Lymedin2010
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I would get this Zeiss & no worries about mercury lamps for fluoresence, as this uses LED based technology. I LOVE my Zeiss lenses for both camera equipment & microscopy, as they are unparalleled.
Price will be 5-6K, but cheaper if you can get a used one.
" -Reflected-light fluorescence -Rapid switching from fluorescence excitation to brightfield illumination -Economical LED concept -Battery pack for operation without a main power supply -Special eyecups eliminate the need for a dark room during a tuberculosis test -Simple to operate -Durable and robust -Tried-and-tested Carl Zeiss optics made from high-quality glass -High-quality materials -Worldwide support from Carl Zeiss"
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Cool. You get great quality with your scope, too Lymedin. Zeiss has a good reputation for optics, camera or otherwise. Can't really go wrong with the brand.
How's the BSK working out so far? Got any long term cultures planned yet?
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A selling point for me is darkfield. I'll have to see if that's compatible with any kits or condensers for that, as well as if that method is compatible with other types of fluorescent stains.
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Dr. MacDonald has suggested Nikon or Olympus as well.
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TNT
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quote:Originally posted by thatdudefromkansas: Once I find a model, my current setup will also be for a sale, and Id let it go for a very reasonable amount for anyone here that wants it. As in beat any price and adjust based on financial situation, just to share the capability to do this yourself.
I know Haley has been wanting a scope for a few years now. I don't know if she got one yet, or not.
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Lymedin2010
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I captured a nice spiro on day 5 of BSK culture & it was very rewarding to have most of the rbc's lyse from freezing and I am very pleased with that method as it eliminates most false spiros.
I have been using Elmer's Glue to seal my slip covers, but it looks like it must be porous & it is drying the slide out over time. I am waiting on the next rounds of trials.
Here is a cheap FM. The mercury bulbs are rather expensive too & burn up rather quickly. One has to keep them on for a few minutes each time you turn them on, as they heat up & give off the proper spectrum of light.
[ 07-10-2016, 07:07 PM: Message edited by: Lymedin2010 ]
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Lymedin2010
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This Chinese import can be had in DF as well & at a fraction of the cost. Even if you need to replace one or 2 of the objective lenses, it may still be worth it.
Will you be doing FITC & fluorescent Bb antibodies or acridine orange?
posted
Nikon makes very good scopes. Just have to find a good deal on them.
And Bluelyme, I'll keep you updated.
As an FYI for you, the scope I have now has both a dry dark field condenser and oil dark field, and I have a 100X oil objective w/ Iris on it as well, so you can use the 100x under darkfield. So really, I have 3 condensers for it. 2 dark, and 1 bright.
There are a few systems to look at. I have considered dropping considerable money on a microscope. One of the higher end models, or even older models. You can find Nikon Microphot systems on eBay and other places online, for example, for between 5,000 and 8,000 dollars.
What I don't want to sacrifice on my next scope is resolution. I am ready to move up a step. My current setup is great, but I want to be able to see the minute details in high resolution at high magnifications, similar to what you would see in legit research labs.
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That's actually pretty good. I am curious how it looks in 4k? Also, I would have to see the setup, as well as actually try it out first, if it is something he makes himself.
I wonder if it is just a camera type attachment that he makes? And not a full microscope system?
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Lymedin2010
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I PM'd you his emails w/description.
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I have zeroed in on a few prospects for microscopes. You can find quite a few serviceable, research grade microscopes from the 80s and 90s, still in use by labs, that are relatively affordable compared to the newest models.
I am seeking out those options, as they will provide the highest quality, highest resolution imaging.
We'll see how they pan out.
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Lymedin2010
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Ha, I thought about posting the link to that one, too. Yes, someone will get a sweet machine. I am watching just to see how high (or low) it goes.
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posted
Thanks for the help in looking for scopes, guys. Input is always good.
I have put the money down for a new one.
I will unveil it when it is appropriate. Got's all kinds of goodies. I bought it used, but everything appears to be in new condition.
High quality lenses. Phase Contrast/Darkfield, DIC, Epi-Fluourescence, and standard Brightfield.
I am actually excited to play around with the DIC. Especially on a high quality. If you don't know what that is, just look it up. Very similar to standard Phase Contrast.
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TNT
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quote:Originally posted by thatdudefromkansas: Thanks for the help in looking for scopes, guys. Input is always good.
I have put the money down for a new one.
I will unveil it when it is appropriate. Got's all kinds of goodies. I bought it used, but everything appears to be in new condition.
High quality lenses. Phase Contrast/Darkfield, DIC, Epi-Fluourescence, and standard Brightfield.
I am actually excited to play around with the DIC. Especially on a high quality. If you don't know what that is, just look it up. Very similar to standard Phase Contrast.
AWESOME!! You're making me drool. Would love to see a pic of it sometime. Can't wait to see some blood with it.
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I'll post a video of the setup and get some BSK samples under it once it's all set up.
Start looking into getting some fluorescent stains and working through that process. Figuring out which, antibody-wise, to start out with.
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Lymedin2010
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Interesting. Also note, Darkfield is preferred method for viewing the nematodes, no fixation or staining required.
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Lymedin2010
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Good stuff, thanks!
Interesting how there is a lot more buzz about Borrelia in the blood nowadays. Sounds like everyone's concerted efforts for recognition has started to pay off.
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Lymedin2010
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E-E-E-E-e-e-e-W-W-W-W-w-w-w-w !!!! THAT, is unmistakable! I'm glad I've never seen anything like that in my blood!!!
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Just took possession of an AO10... excited to re-read this thread and figure out how to get going.
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TNT
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quote:Originally posted by mustardseed2: Just took possession of an AO10... excited to re-read this thread and figure out how to get going.
Way to go, mustardseed2! Looking forward to hearing/seeing what you discover. Don't be afraid to ask questions or post any pics/videos.
Is your 10 equipped with either darkfield or phase contrast?
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I might try a makeshift dark stop at some point for 40x, but I'm more interested in doing staining to check for co-infections.
Looking into how to do Giemsa now.
My first look at my blood cells, I didn't notice anything funny looking, except a couple of them had a large, black, bumpy, V-shaped inclusion. Not sure what those were.
Posts: 71 | From Canada | Registered: May 2016
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posted
So I saw my first potential spirochete tonight.
I haven't read this entire thread, so I'll go back and see if it matched what other people have seen.
It had bulbous tips, and sometimes looked like the string of pearls mentioned earlier. It was oscillating around quite quickly.
Are there any other organisms in the blood that could potentially look like this?
I also saw a couple that were shorter, with a bulb on each end and a short connection in between.
Posts: 71 | From Canada | Registered: May 2016
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bluelyme
Frequent Contributor (1K+ posts)
Member # 47170
posted
Congrats it is scary cool huh?..cant wait to get my own scope
-------------------- Blue Posts: 1539 | From southwest | Registered: Dec 2015
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I did several smears on the first day, and didn't find anything.
On Day 2 I re-checked the smears from Day 1, and still nothing. Then I made new smears, and still nothing.
Then before I went to bed, I did one more smear (this is 8 hours after my previous smear), and all of a sudden I could see a couple of them.
I'm going to draw more blood before bed tonight to see if there's a pattern or if it was coincidence.
edit: I also bought that camera stand from ebay for $20. It's coming from Chine though, so maybe by September I'll be able to post pictures haha.
Posts: 71 | From Canada | Registered: May 2016
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