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

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bluelyme
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   Posted 9/11/2017 3:17 AM (GMT -7)   
/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 3:34 AM (GMT -7)   
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 8:31 AM (GMT -7)   
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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   Posted 9/11/2017 12:11 PM (GMT -7)   
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 11:00 AM (GMT -7)   
Great vid mustard ...wow ...any ideas @ darkfield at 1000x
maybe toots can explain flourencence microscopy a bit

Healrom
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   Posted 9/12/2017 8:53 PM (GMT -7)   
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 9:22 PM (GMT -7)   
Healrom - are those spirochetes in your video that are wiggling???

Healrom
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   Posted 9/12/2017 10:18 PM (GMT -7)   
In which one?

Mustard Seed
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   Posted 9/13/2017 9:48 AM (GMT -7)   
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
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   Posted 9/13/2017 2:31 PM (GMT -7)   
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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   Posted 9/13/2017 4:41 PM (GMT -7)   
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
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   Posted 9/13/2017 5:17 PM (GMT -7)   
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
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   Posted 9/14/2017 1:04 PM (GMT -7)   
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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   Posted 9/14/2017 3:03 PM (GMT -7)   
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
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   Posted 9/14/2017 4:32 PM (GMT -7)   
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
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   Posted 9/14/2017 4:53 PM (GMT -7)   
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
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   Posted 9/14/2017 5:27 PM (GMT -7)   
I have the same flashlight at home ;)

TOOTY
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   Posted 9/14/2017 6:55 PM (GMT -7)   
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
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   Posted 9/16/2017 10:41 AM (GMT -7)   
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
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Total Posts : 231
   Posted 9/18/2017 2:37 PM (GMT -7)   
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

Yellow Cat
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Date Joined Dec 2016
Total Posts : 196
   Posted 9/22/2017 10:59 AM (GMT -7)   
@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.

Mustard Seed
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   Posted 9/26/2017 8:24 AM (GMT -7)   
At what part if the video do you see spirochetes? I had a hard time seeing anything.

Lapis_29
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Date Joined Sep 2017
Total Posts : 442
   Posted 9/26/2017 8:46 AM (GMT -7)   
Mustard Seed said...
At what part if the video do you see spirochetes? I had a hard time seeing anything.


ok, in this video right here

https://www.youtube.com/watch?v=1HUtKungjvE&app=desktop

watch at exactly 44 - 49 seconds, a squiggly line just to the left of the center of the screen turns from a line to a round thingy

this may very well be a spirochete (line) transforming into the cyst (circle) form of borrelia

Lapis_29
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Date Joined Sep 2017
Total Posts : 442
   Posted 9/26/2017 8:57 AM (GMT -7)   
hey microscopy people!

fascinating videos y'all have here!

I would very much be interested in someone doing an experiment. I am having AMAZING results with DMSO treating neuro-lyme. As most knw DMSO is very penetrating. Apply some to your skin and it goes deep into the tissues.

I would be absolutely fascinated if someone here would do a microscopy experiment showing blood, then blood from the same area after being treated by DMSO. I really think this would give some great results.

here is my thread on DMSO with lots of great info.

thanks!

http://www.healingwell.com/community/default.aspx?f=30&m=3914807

Mustard Seed
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Date Joined May 2016
Total Posts : 1152
   Posted 9/26/2017 9:30 AM (GMT -7)   
Lapis_29 said...
Mustard Seed said...
At what part if the video do you see spirochetes? I had a hard time seeing anything.


ok, in this video right here

https://www.youtube.com/watch?v=1HUtKungjvE&app=desktop

watch at exactly 44 - 49 seconds, a squiggly line just to the left of the center of the screen turns from a line to a round thingy

this may very well be a spirochete (line) transforming into the cyst (circle) form of borrelia


Ya clear as day in those videos, I was asking about the videos JohnB2 posted.
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