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» LymeNet Flash » Questions and Discussion » Medical Questions » I want my own Microscope... (Page 2)

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treepatrol
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Thanks James maybe everybody who are checking blood for Bb should do it like this.

ABX on =
Date started=

ABX first of month blood check

ABX end of month blood check

or three sldes at first of month and spirochetes any?

or three slides at end of month and spirochetes any?
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Or the ABX your on=
Date started=

And do the blood check every 10 days = 3 x in 30 days.

That way you catch cycles of Bb and significant abx exposure too.

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Do unto others as you would have them do unto you.
Remember Iam not a Doctor Just someone struggling like you with Tick Borne Diseases.

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James H
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If only it would lend itself to such neat, orderly, and quantifiable analysis!

The problem is that what can be seen in the blood at a given moment does not represent the whole body. In the morning after a good night's rest the blood can appear almost sterile... yet appear totally septic again by that evening. I don't think the infection fluctuates that much, but what is seen in the blood does. The blood is maybe not the focal point of the infection, but it is a convenient thing to examine.

I think daily samples taken at the same time of day could give a general idea of quantity, especially for the same person. There is alot of variance to deal with though.

I'm looking for changes in appearance and condition, and take lots of photos to compare. If membranes look damaged or deformed, or the organisms appear sluggish, that is of interest.

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wrotek
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Starship trooper very nice movie and very nice darkfield, impressed. What a pitty u did not catch moving spiros.

quote:

Wrotek, Take a look at http://biomedx.com/microscopes/training/LB1.html

"CAUSE: protein linkage. Often poor protein digestion. The pancreas may be off. Excess dietary protein, poor assimilation. Eating too much animal protein. Blood too toxic (altered blood pH-zeta potential down) from stress, coffee, cigarettes, meat, etc. Dehydration, not drinking enough water (which by the way, is one of the top undiagnosed causes of many ailments). Eating the wrong foods for the blood type, e.g. wheat consumption by type O's, beef consumption by type A's, etc.

SIGNS: Fatigue, shortness of breath - RBC's cannot carry oxygen; stress on heart. Cold hands/feet - poor circulation."

I drink coffee and i deffinetely notice difference In my hands and feet, they become very cold.

This clothing explenation as a dying blood(thrombosis) and bad diet (stress) is very convincing but i also have seen in Dr Lida Mattman book Stealth Pathogens a photograph where young spirochetes created hemagglutinin that makes erythrocytes rosseting around spirochete.
This is this photo
http://www.borelioza.tivi.pl/photogallery.php?photo=15
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signature of photo is "Young colonies of B.Butgdorferi may make hemagglutinin which causes erythrocytes to rosette around them", any ideas they do it? Easier cross from one cell to another without risk meeting immune system cell ?

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James H
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Interesting pictures. This one looks alot like what Starship Trooper was showing us:



That picture you linked is definitely not normal.

It is just that some people will point to any kind of normal appearance or normal degradation of an old sample and use it to scare people so they buy products. I am sure you know what I mean.

Eastern Europe is WAY ahead of the US in studying this desease and looking for practical ways to deal with it. If anyone finds a cure, it will be them, using 50 year old equipment, not our 'head in the sand' US CDC geniuses.

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wrotek
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This pictured You showed is multiple sclerosis spirochete also from Dr Lida Mattman Book-Stealth Pathogens.
Dr Lida Mattman is not selling anything, she is microbiologist nominated to nobel prize in medicine

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James H
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I was not referring to Dr. Lida Mattman in that way... she is of the kind I greatly admire. [Smile]
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david1097
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OK guys... for your viewing (dis) pleasure try these links:

http://www.cdfound.to.it/html/atlas.htm#atlas
http://www.cvm.okstate.edu/~users/jcfox/htdocs/clinpara/clinpara.htm
http://www.cvm.okstate.edu/instruction/kocan/vpar5333/deerpar.htm

And James, what you have seen in the RBC's has been looked in some detail and PCR'd to confirm that they are some type of bacteria.... even in "healthy" people. It looks like it has been reported several times and once in the journal "Nature" so it appears to be able stand the assualt of peer naysayers...... Still it looks like nobody listens (or does not want the added complication in their lives... or maybe know something that everyone else does not.)


Heres the link to recent paper with references. They apparently stumbled across this using dark field trying to spot spirocettes in some degerenative brain disporders (sounds familiar?)

http://jcm.asm.org/cgi/reprint/40/12/4771

Let me know what you think.

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James H
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Thanks, David. That last link was interesting. It seems they isolated Pseudomonas maltophilia from some people's blood.

From the pictures I have seen elsewhere of Pseudomonas maltophilia I don't think that particular bacteria is the same as the darkfield forms I am seeing, though some of the rod forms look similar.

It does show that they CAN identify an unknown bacterial organism in blood fairly easily if they want to, and that blood is anything but sterile.

These strange looking forms have been noticed as early as 80 or 90 years ago, and they really have not been studied much in all that time.

I don't automatically assume that everything seen is a pathogen, they may just be interesting 'things'.

How about this guy?

Image hosted by Photobucket.com

[ 18. January 2006, 01:03 PM: Message edited by: James H ]

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caat
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Wow. I missed a lot this week. So much great information! Thanks James and thanks David for posting that atlas etc.

Congradulations on that home made darkfeild filter Starship Trooper, that's impressive! Do you have a photo of it? I can't understand it from the diagram but might understand a photo. Where are the glass lenses underneath the stage from? Or is that the condenser and you tape cardboard under the stage? Maybe it's not that simple...

I still haven't ordered stain yet, I think my liver is rebeling and I can only do about 3 things per day right now. I feel like I drank a case of whiskey last night...(It's OK though- I'm ramping down and ending this protocol and should start feeling a little better in several days.)

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millymollymandy
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What a fascinating post! I'm going to have to come back and read it properly later on. I have just looked at a couple of the sites that were linked and somehow I really don't fancy my breakfast now! Yuk!!!
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david1097
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here's another link that might be of interest
http://www.diaglab.vet.cornell.edu/clinpath/modules/rbcmorph/rbcmorph.htm

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Starship Trooper
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OK, Here's a detailed explanation of how I did it.

First a little about the microscope. You will notice from the pics that the objective is quite big. In fact the whole thing was bigger than I expected. I guess that the bigger the lenses, the better the pictures but the size shown here is a standard in microscopes called the DIN standard.

This means that you can go and buy an objective from any local shop and it will fit.
This is the objective I used.

 -

Just under my finger is what you place the slide on and that is called the "stage". There is adjustment underneath the scope so that you can move the slide around without touching it yourself.

Under the stage is the condenser. This is responsible for focusing the light onto the slide you are looking at. It is important to have an adjustment on this for the dark field, but a little hard to explain why..

This is the condenser under the stage. I am pointing to a filter holder where we will be putting our filter. This just slides out.

 -


Sliding out the filter holder, you see this.
 -

In fact, this is a bright field condenser with a filter holder. What I am going to explain below uses this condenser. I have asked about a dark field condenser and they are available. Because this Microscope is standardised, I am able to just buy one off the shelf. This could be a decision if you decide to get your own.

Here is the condenser removed. Note that I have inserted the filter in the holder in this photo.

 -

OK, that said, lets get on to making the filter. The filter is a device to block the light from going into the objective. The condenser focuses a ring of light onto the slide all into one point, so the slide is well lit.

When the objective sees it, the only light it sees is from the objects on the slide. This is why you see light objects on a dark background.

These were the disks I made to produce the hole in the light.

 -

Next I laminated them and cut them out so that they would fit into the filter holder.

 -

The disks need to be central within the laminate and they need to be as round as you can cut them.

Place them in the filter holder and look at a slide through the scope. I found it best to use a 10x objective and a 10x eyepiece lens. Yep that is just 100x total. This is to get your dark field working.

You will have to alter the adjustment for the focus of the condenser so that you see bright blood cells on a black background. If you do not see this, change the disk for a larger one and repeat until you do.

Once you have the darkfield sorted at 100x, connect your camera or video camera to the eye piece as shown and zoom to about 8 times. This takes a little setting up as you have to the lenses straight.

 -

You may have to slightly refocus on the microscope for the camera to see it better.

Once you have got this far, looking is all there is left to do. Instead of looking through hte eyepiece, you can now look on the screen of your camera. Move the slide about using only the adjustment for the stage on the scope.

One thing that crossed my mind was that my pics are the classic spirochete form, where by others are rolled up into cysts. This may be because I am on doxy and they are not. Either way, another interesting fact is that my blood was taken late in the evening. I don't think they are in the blood so much in the morning. One ref to back this up is at http://www.canlyme.com/lymeconf04.html

Skip down to the part on Nick Harris and lyme testing.

Another interesting fact about my logs is that I have a +ve PCR - first time. Don't know if this was good luck or that there is something in it, but out of the two smears that I have done, I have seen Keet looking bodies in both of them.

If you have any further questions, I will gladly answer them.

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Starship Trooper
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Microscope rentals - ships world wide!

http://www.microscopestore.com/microscope-rentals.asp

Edited - Actually saying that they do look a bit pricey

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caat
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Starship, that was very clear. Thank you for taking the time to make a DIY lesson! I've saved this web page to look at later if I get a scope with a condenser. And it helps to know what condenser to look for (w/filter holder).

[ 24. January 2006, 04:19 PM: Message edited by: caat ]

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caat
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I'm starting to look to order giemsa stain and what others might be useful later.

One of the manuals says to dilute giemsa with ph buffered water.

Does anyone know if I can just use baking soda or vinegar to buffer with or would those interact with the stain? Do I need something special? maybe citric acid & ???

And if giemsa has any other name? So far all the ones that carry it seem to be professional's companies. I know I had a problem ordering orchid sowing materials before from these- many want a title or company to send to.

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Starship Trooper
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quote:
Originally posted by caat:
I'm starting to look to order giemsa stain and what others might be useful later.

One of the manuals says to dilute giemsa with ph buffered water.

Does anyone know if I can just use baking soda or vinegar to buffer with or would those interact with the stain? Do I need something special? maybe citric acid & ???

And if giemsa has any other name? So far all the ones that carry it seem to be professional's companies. I know I had a problem ordering orchid sowing materials before from these- many want a title or company to send to.

Hi Caat,

I ordered mine in a pre made solution from www.ralamb.net Their part number was LAMB/120-D. I am not sure if they do a ready-made solution in your part of the world, but they will sell you the powder to make it up yourself. It might be worth asking them for this part number, as they are the same company just a different country.

The buffer solution can be deionised water. You should be able to get this in most super markets as people use in irons to prevent them from scaling up. Alternatively use distilled water. Mix 9 parts stain to 1 part water.

The problems I have run into (not big ones) is that the stain seems to have microscopic hair like things in it and other debris. It needs to be filtered just before it is put on the slide.

Another thing you need to have is ethanol to stop the RBC's from bursting when the stain is applied. I tried it with some vodka for 5 minutes and it seemed to be OK! - Don't drink it afterwards though...

When I get a better procedure together I will post it here. Also, just in case people don't know how a smear is done, I can probably do a small example video.

ST.

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caat
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ST,

Thanks!

I think distilled water has a ph of 5.something. From what I've read the water should be buffered to between pH 6.8 & 7.2 for diluting giemsa.

They usually use phosphates as a buffer; a mix of sodium phosphate and potassium phosphate, although it seems you could use one or the other. I found no mention of baking soda, although it was mentioned as a buffer for other stains. The phosphate mix goes for around $6 in the US.

Of course we probley don't need to be as particular...

I'm going to try Wright's stain first. Haven't read the manual on it yet.

Thanks for the tip about vodka!! I was going to buy wood alcohol in the hardware store today, but wasn't sure if it should be straight methenol or if it could be what they sold- methenol ket-something.

Vodka is a lot less toxic and I can dispose of any left over to some of my neighbors who will be very happy about that. I will just have to listen to them fight later... LOL

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Starship Trooper
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OK, still trying slightly different techniques for the staining, but thought you might like to see some results.. Not absolutely sure what I am looking at!

Caat, pure water has a ph of 7. If you use the wright stain, I believe it has a fixer in it so you can skip the ethanol bit. 90% ethanol is pretty easy to get hold of though.

I have also found that the pictures come out better if the eyepiece on the scope is removed.

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Getting Better
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We've seen videos of the spirochetes on the Canadmian Lyme site. They were actually very frightening to see, burrowing into the B cells. So I wonder what kind of technology was used in finding those and I wonder why tests for Lyme aren't finding the spirochetes on this kind of technology.

I saw my blood many times on the bradford microscope. I have a severe case of Lyme, reinforced by positive Western Blot. The spirochete could not be seen on this microscope. All we saw were signs of infected B cells, and the cyst form -- both indirect speculations.

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Jeff

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Dave6002
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I think the video on that website was in vitro culture not from in vivo.

A European scientist suggest that looking for "apple" instead of "snake" when you check your blood under a microscope.

Because the spirochetes have become granuated. Still a puzzle that why PCR cannot detect the granuated Bbs, he suggested that the granuated Bbs have RNA not DNA. Of course this is a wild thinking. I doubt it.

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ChrisBtheLymie
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You might want to check this excellent new site out;
http://www.lyme-diagnosis.org.uk/

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burnbitter
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hi,
Just thought I'd mention that I have a crappy little microscope.. So I wouldn't be able to see the bb anyway. But you can look at other things like yeast.
But I was using it the other day and I have so many floaters in my eyes that I can't really even see anything. I'm assuming that since this is a common lyme symptom that others will have the same problem. So before you blow a bunch of money on a microscope, you might want to check out the number of floaters in your eyes.

I did the biology thing back in college and the floaters weren't there then... so it was a bit of a surprise.

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Thereminator
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Ive been watching the PBS special Malaria:Fever Wars and I noticed that the scientist at Oxford quick-dry their blood samples with hair dryers.I assume the parasites are much easier to identify,because of their size.
I am excited about this thread
and the Knock-Out clinical trials thread and I hope they stay active!
Im going to keep mentioning the fact that we dont compile personal treatment data(like on remedyfind.com)untill the administrators of this site create a format or easy links ...for that reason!
[Smile] Alan

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Charter member of the ~ Delux Toasting Club ~
Our Moto:
"Take No Prisoners"

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