Microscopy for dummies

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gfields
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   Posted 9/13/2017 5:12 PM (GMT -7)   
Calling all microscopy experts...

I am looking for a darkfield microscope that I can hunt for spirochetes with. I'd like to be able to display the output of the microscope on a Win 7 computer. I'd like to keep it around 600 or less. What kind of scope could I buy to fit this bill?

If there's one on ebay that would do this, could you send me a link, and I'll pick it up?

I just don't really know enough about scopes to be able to make this call, but I would like to give it a shot. Thanks for your help!!!

gfields
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   Posted 9/13/2017 5:18 PM (GMT -7)   
Would one like this work well for spirochetes?
http://www.microscopenet.com/omax-40x2000x-darkfield-trinocular-compound-microscope-with-digital-camera-p-10243.html?gclid=CjwKEAiAvs7CBRC24rao6bGCoiASJABaCt5DpGWSgZWPThqIzpZd7NIpz8gYP9-mpvnmXC-6TS9pqRoCVrnw_wcB

Mustard Seed
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   Posted 9/13/2017 6:39 PM (GMT -7)   
Hey gfields,

When doing darkfield (if that's what you're looking at doing), you always want the N.A of the objective lens to be less than that of the condenser.

This means that usually for 1000x darkfield, you need an objective lens with an adjustable iris (to lower the N.A) or a funnel stop that you manually insert into the objective (again to lower N.A). It's unclear whether the 100x objective lens with this kit includes either of those two, but I suspect not.

I'm also not sure if that bulb is strong enough for 1000x darkfield, but someone in the microscopy thread might know.

To be honest, it seems a little expensive for what you get, given that a lot of times these Omax or Amscope cameras really don't work well.

Check out this thread I made a while ago on basic microscope stuff:

www.healingwell.com/community/default.aspx?f=30&m=3753190

Mustard Seed
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   Posted 9/13/2017 6:48 PM (GMT -7)   
Just to add onto my last post, look how this microscope specifically comes with a second 100x objective that has an adjustable iris:

www.amscope.com/compound-microscopes/darkfield-trinocular-biological-compound-microscope-40x-2000x-1.html

If you're looking to do darkfield at 100x, that's something you'll probably want to look for. Also, it specificially lists it as a "plan achromatic" lens, which doesn't blur the edges of the image like a regular achromatic lens (this isn't something that's super important, usually costing more $, but it's a good bonus).

gfields
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   Posted 9/13/2017 7:00 PM (GMT -7)   
Oh yes, that's a wonderful post. I was just reading that a little while ago. My brain petered out half way through though. But thanks for putting that together. It's extremely helpful.

I'll try reading that post a bit more in depth tomorrow.

Is there one on ebay or something that you'd recommend as a better one? I'm not super knowledgeable about which ones are good and which ones are bad.

I'd like to get a spinal tap and analyze CSF under the microscope. I have pretty severe neurolyme. I'd be interested to see if I could spot a spirochete in the CSF fluid if the doctor would let me take a sample home. Then, if I could hook it up to a computer, I could take screen captures, or videos of what I found, and show them to my LLMD. That would be pretty cool.

...

Oh, sorry, your other post came in while I was writing this one. OK, I'll check this one out. It's still in my price range. I wonder if you can hook it up to a laptop. I'll dig in further. Thanks so much!!!

Post Edited (gfields) : 9/13/2017 8:03:16 PM (GMT-6)


Mustard Seed
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   Posted 9/13/2017 7:15 PM (GMT -7)   
My best advice would be to not tell any doctor that you're looking for spirochetes under the microscope; they tend to immediately devalue anything you say from that point forward.

You're just as likely to find spirochetes in your blood as your CSF I'd reckon. I'll look on eBay a bit later.

The way I got mine was to join a microscope forum like microbehunter, and see if anyone's selling. That way you're buying from someone who knows microscopes and probably kept good care of it.

Or just look for Amscope or Omax, I've seen some nice looking videos from some of those. Maybe just buy your own camera, as I've never heard a positive review of an am scope USB camera. I would buy a scope first and worry about the camera later once you've honed your technique.

Post Edited (Mustard Seed) : 9/13/2017 8:18:08 PM (GMT-6)


Healrom
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   Posted 9/13/2017 7:32 PM (GMT -7)   
Hi, I would advise you an AMscope.
A halogen version, not led. With 10 and 20x eye piece (objective)

And either you buy a model that is sold with an OIL darkfield condenser. Or you buy a model without and you buy a darkfield oil condenser separately.

It's very important to take an OIL condenser, and not DRY.

You will only be able to use the 40x objective with darkfield, but it's really enough to search and see spirochetes.

So it will be a 2000X compound microscope with halogen illumination.

Then if later you want to do darkfield with 100x objective, you will have to buy an iris 100x objective, and maybe customize the lighting part.

You can take a binocular (and you will have to put a camera at one of the eyepieces) or a trinocular, dans then you can put a camera at the third tube. If you want to use a photo camera with an adapter (but they are expensive), I would advise you a trinocular.

Other than that, you can use a webcam taped to an eyepiece to film videos. I'm using a Logitech one.

gfields
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   Posted 9/15/2017 2:31 PM (GMT -7)   
So it sounds like you guys are both recommending that I go with this one:
http://www.amscope.com/compound-microscopes/darkfield-trinocular-biological-compound-microscope-40x-2000x-1.html

Psilociraptor
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   Posted 9/15/2017 3:21 PM (GMT -7)   
I have an Omax with a camera. The camera is actually pretty good with healthy expectations. When imaging something like a spirochete you are literally observing the thinnest visible organism on earth. It can be hard enough to see directly, much less with a camera. The camera will never look as good as directly looking through the microscope either. As for the scope itself I personally love it. I hear a lot of poo pooing about omax but it honestly works great. The one thing I have not tried is the 1000x setup however so I can't comment on that. Omax sells a set up specifically for that but I can't say how good it is.

gfields
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Date Joined Oct 2015
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   Posted 9/15/2017 4:03 PM (GMT -7)   
Is this the omax you own?
http://www.microscopenet.com/omax-40x2000x-darkfield-trinocular-compound-microscope-with-digital-camera-p-10243.html?gclid=CjwKEAiAvs7CBRC24rao6bGCoiASJABaCt5DpGWSgZWPThqIzpZd7NIpz8gYP9-mpvnmXC-6TS9pqRoCVrnw_wcB

I was wondering about something. With a microscope, in theory, you could experiment with your own personal spirochetes, to see if your chosen method of treatment is effective in killing spirochetes, couldn't you?

Could you dilute abx in water, to what you think might be a level that would be exposed on a cellular level, and have one drop of blood on a glass film as a constant, and another drop of blood on a glass film where you introduce a drop of your diluted treatment, and see if the spirochetes die on the diluted treatment glass film, and they live on the constant glass film.

I wonder if you could do the same thing with the Spooky II. Have a constant glass film and a glass film with spirochetes treated on a "spirochete kill" setting, and see if they, in fact, die on that setting.

Has anyone experimented with this type of thing?

Mustard Seed
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Date Joined May 2016
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   Posted 9/15/2017 5:37 PM (GMT -7)   
gfields said...
So it sounds like you guys are both recommending that I go with this one:
http://www.amscope.com/compound-microscopes/darkfield-trinocular-biological-compound-microscope-40x-2000x-1.html


This microscope would certainly have everything you need. You could look for Borrelia in darkfield and co-infections with stains in brightfield. The only thing I'm not sure about is if a 20 watt bulb is bright enough for 100x darkfield. You'd assume that if they're selling it with the 100x iris objective it would be, but 20 watt seems a little dim IMO, but I've never experimented with one.

gfields
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Date Joined Oct 2015
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   Posted 9/15/2017 7:46 PM (GMT -7)   
Maybe I could ask them if I could upgrade to a brighter bulb. Would a 40 watt bulb be more appropriate perhaps?

Which microscope are you using currently?

Post Edited (gfields) : 9/15/2017 8:53:31 PM (GMT-6)


gfields
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   Posted 9/15/2017 7:59 PM (GMT -7)   
microbehunter is a pretty cool site. I found a thread about replacing bulbs:
http://www.microbehunter.com/microscopy-forum/viewtopic.php?f=5&t=5122

I'm no electrician, but I bet I could find a youtube video online about this and try to follow the steps if I come to the conclusion that the bulb brightness isn't going to cut it.

Healrom
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Date Joined Nov 2016
Total Posts : 133
   Posted 9/16/2017 8:22 PM (GMT -7)   
The amscope seems good. I think that there wasn't complete kits with 100x iris objectives at this price before.

For the light bulb power I'm not sure that you can change it like that. Because the electronic circuit may encounter problems after that.

I think that the best is to ask to the seller if the intensity of the light souce will be enough in 100x darkfield if you want to be really sure.

I have an amscope, but a second hand 100x ojective, and a custom lighting system, so I can't tell you.

Healrom
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Date Joined Nov 2016
Total Posts : 133
   Posted 9/16/2017 8:25 PM (GMT -7)   
Also the 100x objective of amscope is not very good. But the one I have is a basic one. This one is a plan objective, so it should be better.

Mustard Seed
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Date Joined May 2016
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   Posted 9/17/2017 6:46 AM (GMT -7)   
gfields said...
Maybe I could ask them if I could upgrade to a brighter bulb. Would a 40 watt bulb be more appropriate perhaps?

Which microscope are you using currently?


I use an American Optical 10. It's an older microscope but it's in great condition. The reburbisher I bought it from let me know that my 18 watt illuminator is not enough for 1000x dark field. The American Optical darkfield scopes were sold with 100 watt illuminators back in the day.

gfields
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Date Joined Oct 2015
Total Posts : 867
   Posted 9/17/2017 7:29 AM (GMT -7)   
Something like this?
http://www.ebay.com/itm/AMERICAN-OPTICAL-FULLY-EQUIPPED-MICROSTAR-2070-VERTICAL-FLUORESCENCE-MICROSCOPE-/292192313155?hash=item440804df43:g:W8EAAOSw4dxZc4aj

Mustard Seed
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   Posted 9/17/2017 8:31 AM (GMT -7)   
It's hard to know the quality of stuff you buy on eBay unless you personally know the seller. A bit of a risk when you're spending big money like that.

If I had to buy again without knowing any sellers, I'd probably buy a new Amscope with a 100x objective with adjustable iris, and a darkfield oil objective. Start with brightfield + quick dip veterinary stains for co-infections, and 1000x darkfield for spirochetes. You can do fancier stuff later.

Healrom
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Date Joined Nov 2016
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   Posted 9/17/2017 10:11 AM (GMT -7)   
Yes I agree with that.

You can even begin the darkfield with 40 X objective, I did it for years and was happy with it. The setup is more simple than with 100x.

gfields
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Date Joined Oct 2015
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   Posted 9/25/2017 11:35 AM (GMT -7)   
I took the plunge and ordered the Amscope. Should be interesting.

gfields
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Date Joined Oct 2015
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   Posted 9/30/2017 10:35 AM (GMT -7)   
This microscope pretty much does the same thing as the old 1980s microscope that I currently have. Same exact magnification lenses. I guess there's not a lot of sense keeping it, if it does the same thing as my current microscope. Maybe I'll try to exchange it for one with a bit higher magnification. Not sure. It also did not come with the darkfield piece that it said that it would come with.

Do we need to use a special stain to see spirochetes? Is there something that we need to put in the sample to keep them alive?

What I did see very clearly with this microscope is the enormous amount of eye floaters I have in my eye. That's really quite frightening. They are in the shape the spirochetes, and they have little "blebs" all over them. Little round things. I wonder if I have tons of spirochetes hiding in the gel in my eyes. Maybe they go there to escape the abx?

In both microscopes, I could see red blood cells, but I don't think the magnification is quite high enough to see them well. They both has 1000x magnification as the highest level, but I think 1500x magnification might be better to really see them up close.

Mustard Seed
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Date Joined May 2016
Total Posts : 1153
   Posted 9/30/2017 3:39 PM (GMT -7)   
gfields said...
This microscope pretty much does the same thing as the old 1980s microscope that I currently have. Same exact magnification lenses. I guess there's not a lot of sense keeping it, if it does the same thing as my current microscope. Maybe I'll try to exchange it for one with a bit higher magnification. Not sure. It also did not come with the darkfield piece that it said that it would come with.

Do we need to use a special stain to see spirochetes? Is there something that we need to put in the sample to keep them alive?

What I did see very clearly with this microscope is the enormous amount of eye floaters I have in my eye. That's really quite frightening. They are in the shape the spirochetes, and they have little "blebs" all over them. Little round things. I wonder if I have tons of spirochetes hiding in the gel in my eyes. Maybe they go there to escape the abx?

In both microscopes, I could see red blood cells, but I don't think the magnification is quite high enough to see them well. They both has 1000x magnification as the highest level, but I think 1500x magnification might be better to really see them up close.


Is this the microscope you bought?

www.amscope.com/compound-microscopes/darkfield-trinocular-biological-compound-microscope-40x-2000x-1.html

If so, it should be able to do both brightfield and darkfield at 1000x. It also come with 20x eyepieces, meaning you can do 2000x magnification (100x objective + 20x eyepiece), although the resolution will be a lot less (higher magnification eyepieces are akin to using digital zoom on a computer vs true magnification that comes through the objectives).

Did your old microscope do darkfield at 1000x? The darkfield comes through the darkfield condenser, not through the objectives. The important pieces in this set are the addition of an oil darkfield condenser (aside from the abbe condenser used for brightfield) and an 100x objective with an adjustable iris. If there's stuff listed on the page that you didn't receive I'd get a hold of the manufacturer and let them know there was an error.

As far as exchanging for a higher magnification, 100x objective is as high as you'll get. Don't forget when using the 100x objective, you need to use it with immersion oil (both on the objective and condenser) or your image will be blurry. You don't need any stain to see spirochetes, especially in darkfield. Check out the lower part of this post if you want information on using immersion oil and how to using a cover slip to smear your slide:

www.healingwell.com/community/default.aspx?f=30&m=3753190

gfields
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   Posted 10/2/2017 1:54 PM (GMT -7)   
After taking it all apart, and putting it back into the styrofoam box, tapping it up, ect. I turned the box over, and then I found the darkfield piece and another optical piece tapped to the outside of the box, in this styrofoam nook.

Then I re-unpacked everything that I had just packed up, and tried the darkfield condenser and the other optical piece. With the 40x optical piece, I could see blood cells. It was like looking at them from far away. Then I tried the 100x and I could see them but they looked like little red blobs. I tried with the 1000x piece, and I couldn't really make out anything. There was some sort of texture, but it didn't really look like anything. Then I tried the optical piece that was taped to the outside of the styrofoam along with the darkfield condenser, and again, it just looked like texture. I couldn't see anything.

It was a frustrating experience, all-in-all. Amscope should really provide documentation about how to use this equipment. The microscope comes with at least 15 pieces that you need to figure out how to put together. There's no instructions except a little pamphlet telling you to search youtube to find videos. When you do that, you find there's a gazillion different Amscope videos, and you kind of have to piece together the information yourself.

Then I took out my old microscope from the 80s and I was able to see exactly the same stuff I could see with the Amscope microscope. I guess I could have been using that silver optical piece incorrectly, because I could only see textures. Maybe I should have used the oil like you were saying. There was a little bottle of yellowish oil that came with the microscope, but I wasn't sure if that was for putting on the slides or if it was for oiling the gears on the microscope. There wasn't any label on the bottle telling you what it was for, or what it was.

My old school microscope doesn't have a darkfield condenser, but I think I can make one by simple drawing a black circle onto a piece of clear plastic. I saw a youtube video on that. Isn't that all the darkfield condenser is? A black circle?

...

I just looked at the Amscope video again, and they say you need to put that oil on either the condensor or on the slide. I think I remember seeing that the first time, but I didn't understand what the point of the oil was. It seemed strange to put oil on the outside of the slide, and then smoosh the eyepiece into the oil. I wasn't sure if there was a glass slip covering the sample or if you were putting the oil directly into the sample and mooshing it into the blood. The video also said you could put the oil on the darkfield condensor, which is maybe what I should have done.

Do you think if I used the oil, I would have been able to see more than just texture with the 1000x eye piece? Is the oil the key to being able to see spirochetes?

The only eye piece that I could see anything with was the 40x.

This stinks, because I just sent the microscope back to the company. I became very frustrated with it, but maybe I should have waited and played with it some more.

....

I was also wondering if you need to use an anti-colagulant solution for the blood so it doesn't stick together. It seemed like after about one minute, the blood coalgulates and sticks together.

....

This guy kind of shows you how to make a darkfield microscope cheaply. Not sure if this is the same as the oil immersion:
https://www.youtube.com/watch?v=taKfysZ-LNY

Post Edited (gfields) : 10/2/2017 3:29:09 PM (GMT-6)


TOOTY
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Date Joined Apr 2014
Total Posts : 231
   Posted 10/2/2017 3:24 PM (GMT -7)   
gfields said...
I just looked at the Amscope video again, and they say you need to put that oil on either the condensor or on the slide. I think I remember seeing that the first time, but I didn't understand what the point of the oil was. It seemed strange to put oil on the outside of the slide, and then smoosh the eyepiece into the oil. I wasn't sure if there was a glass slip covering the sample or if you were putting the oil directly into the sample and mooshing it into the blood. The video also said you could put the oil on the darkfield condensor, which is maybe what I should have done.

Do you think if I used the oil, I would have been able to see more than just texture with the 1000x eye piece? Is the oil the key to being able to see spirochetes?

The only eye piece that I could see anything with was the 40x.

This stinks, because I just sent the microscope back to the company. I became very frustrated with it, but maybe I should have waited and played with it some more.


This guy kind of shows you how to make a darkfield microscope cheaply. Not sure if this is the same as the oil immersion:
/www.youtube.com/watch?v=taKfysZ-LNY



YES, to see anything at 1000x in darkfield and brightfield, you MUST use oil with the 100x oil objective. That means you put a drop of that immersion oil on the slide and carefully lower your 100x oil objective down into it. Once it touches the oil then look through the eyepieces until you bring the image into focus.

When using an oil darkfield condenser, you must use a drop of oil between the top of the condenser lens and the bottom of the specimen slide.

If you don't use immersion oil with an 100x oil objective, you will only see "texture." Especially when it comes to darkfield. But, DO NOT use oil with anything but your oil objectives (and oil darkfield condenser), since that will ruin non-oil lenses.

That video shows a non-oil darkfield hack. But, if you are careful, I think you can use oil on a regular abbe condenser lens and not ruin the condenser lens. I think "bluelyme" has done it, but you have to be careful not to get oil down into the condenser. But, unless you have a 100x oil objective with an adjustable iris or a funnel-stop, you will not be able to do 1000x darkfield.

gfields
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Date Joined Oct 2015
Total Posts : 867
   Posted 10/2/2017 4:33 PM (GMT -7)   
Hmmm. Too bad I mailed it back today. It cost me 70 dollars to mail it too. Maybe I should re-buy it? I guess once I get the refund, I'll only be losing 70 dollars because there is free shipping.

Or maybe I should try the non-oil darkfield hack, and the abbe condenser lens. What is an abbe condenser lens?

So does the 100x lens turn into a 1000x lens when you add the oil to it?
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