Protein and Collagen, what scope can see it?

[Dr.Mary]

Protein and collagen is what I need to see, is this an electrical microscope I need for this?

Please direct me, I’m still getting educated regarding what all I’m to look for so thanks to those that have been so kind to help with the questions, hope they are not all over the place. I wish to purchase the microscope once, so if there is an older one that can be made to “see” what I’m wishing to see (protein/collagen/blood disorders) then this would be fine.

I honestly do not know and need the experts here to help, you folks seem to know so much more than most!


BTW, it’s in the plan to paint some of the things you nice people have posted, I’m a hobby artist and love the critters that some of you have posted pictures of, especially the things from under the ocean they are great! My biology teacher (years ago) told me I should do the art in biology books. I have learning problems but learned if I could draw what I saw then I learned it - it worked! Aced the test every time just by drawing what was seen. Heads up for those still in school

[zzffnn]

Collagen: polarization should reveal some. Phase contrast may reveal collagen too, along with large amount of protein aggregates.

[Dr.Mary]

Collagen: polarization should reveal some. Phase contrast may reveal collagen too, along with large amount of protein aggregates.

That’s a bit above my head, thank you but I do not understand what you are saying. I’m too new to this.

Let’s try this, for an electron microscope (which is what I think I’ll prolly need) do I want the transmission TEM- EM or the Scanning SEM-EM? And why?

[zzffnn]

TEM more likely; I am not positive. Honestly though, TEM or SEM is not for regular hobbyists. I don’t mean to be rude, however if you need to ask, you probably won’t be able to run TEM/SEM without professional help. I certainly cannot run it myself.

Phase contrast and polarization are much easier to run.

[Dr.Mary]

TEM more likely; I am not positive. Honestly though, TEM or SEM is not for regular hobbyists. I don’t mean to be rude, however if you need to ask, you probably won’t be able to run TEM/SEM without professional help. I certainly cannot run it myself.

Phase contrast and polarization are much easier to run.

Thank you, and your not rude at all I appreciate the honesty. I am realizing I’m out of my comfort zone asking these things but there is a goal in mind here. Didn’t realize there was a lot more than setting it up and looking through it but from some of the videos I’ve seen this makes it easier for me to understand what your telling me.

I still haven’t given up yet, and will look at the TEM you suggested. If it’s just setting it up then ok, if it’s years of study on setting it up then that I don’t have. I’m educated, but don’t have the time In years if that is what is required.

[PeteM]

Proteins can often be tagged and identified with fluorochromes and fluorescence microscopes.

As already said, collagen is commonly birefringent and may show up in simple polarization.

It could be that neither are likely to help you all that much in self-help medical diagnosis.

Owning and running most electron microscopes is like buying a rare Jaguar sports car with electrical and lighting systems provided by Lucas - a company known as the "Prince of Darkness" for the difficulties owners faced in keeping its systems running. Competent electron microscopes are typically complex, expensive to buy and/or keep running, and require sample preparation beyond the means and abilities of most of us.

As a very rough guesstimate --- for every 1,000,000 people who have looked through a microscope, 100,000 hobbyists could easily manage simple polarization. There might be as many as 1000 doing fluorescence microscopy outside of academia and research and just 1 or 2 hobbyists with a running electron microscope at home in their garage.

[Alexander]

Without professional training you will have no chance to properly set up and operate a TEM, even after years of learning and experimentation.

A TEM looks like that:



It costs a 6-digit number of bucks. Just the ultra-microtome you need to cut the specimen into slices of 0.1 micrometers will cost more than most light-microscopes.

Are you prepared to store and handle liquid ethane to shock-freeze your samples at -210 degrees Fahrenheit?


Back to matters: What is your exact need? Do you want to localize proteins and collagen within samples or do you want to study their structure? Fluorescence microscopy allows to identify things like proteins in a sample but does not reveal their structure. It is not an easy thing to do as well. The fluorescence markers needed are very expensive, dangerously carcinogenic and not available for hobbyists. The needed microscope is within a generous hobbyists budget. If you want to study the structure of proteins there is no way around the electron microscope.

[Phill Brown]

It seems likely that the budget and time committed to cover so much ground that has already been covered would impact quality of life in a domestic setting.
It would help to be part of a team who already have years invested in the knowledge and skills to break new ground.
I am very much in favour of investing in quality of life and health is wealth.
SEM has so many limitations including what else you could be doing. Only a serving suggestion.

[Wes]

Hi Dr. Mary,

If you can elaborate a bit further on your goal and expectations we might be able to provide more specific advice. A pathologist would be able to make great use of cheap (under 1000 USD) microscope, provided a histology technician supplies him/her with nicely stained tissue sections (common practice in hospitals). A biologist studying the molecular and/or cell biology of collagen might need a confocal microscope which is out of reach (unless you are rich, I certainly am not). A structural biologist might need a cryoelectron microscope or an X-ray crystallography device.

So to get a productive reply and start looking at collagen and protein please let us know what exactly is it that you want to achieve here. There might be a much better suited technique that has nothing to do with microscopy to provide you with the data you desire.

Best
Wes

[Greg Howald]

I would recommend that you return to the first suggestion. Get on you tube with Oliver Kim and learn about polarization and maybe oblique lighting. Try the stuff that costs less than 20 bucks first. Polarizing sheets, 1 1/4 inch hole punches and opaque black paper are all available on Amazon. So learn, make and have fun.
Greg

[SuiGenerisBrewing]

What do you mean by "need to see". If you want to detect collagen in tissue sections, you can stain for it with Movat's pentachrome and then image on pretty much any microscope. If you want to see molecular structuring, you need about a million dollars for a SEM, and someone who knows how to run one.

[Tom Jones]

No offense, but I think you need to go back and reassess what you want to do, and why. You say you want to seen protein and collagen, but some pretty simple things like phase contrast and polarization are a bit over your head.

To be brutally honest, until you can be a lot more specific about your quest, this whole enterprise is way over your head.

You have a HUGE amount of learning to do before you you should even think about acquiring any microscope to look for protein and collagen. You haven't even mentioned what kinds you want to visualize, where they are, or why. It doesn't sound like you know. You need more background education than any forum or group of forums can ever provide.

And as for scanning or transmission electron microscopes, I really doubt you have the time, money, and patience to to get one set up and running. The maintenance contract alone would break most of us. I've done clinical microscopy for 40 years and would love to have and electron microscope to play with. I did TEM microscopy when I was in college, and was actually playing with an SEM a couple of months ago at the university I attended. I'll tell you straight up front, it's way too expensive, and way too complex for most amateurs. In fact, most research facilities have dedicated TEM/SEM microscopists and core labs to do the microscopy, because it's too expensive and too tedious generally for the researchers and grad students to learn to do well enough to get what they need. It's not a good use of their time and money, so they share the costs with other researchers and concentrate the necessary talents in dedicated microscopists. I've met one guy who has an SEM in his basement, but he was trained as an electrical engineer so he can maintain his old scope by himself, and build replacement parts as necessary. He uses it for photography and some contract research.

Why do I think what you really want is to look at are protein and collagen dietary supplements?

[Zeiss mensch]

hey hi
You ask a vague question. To get a usable answer involves more specific qualifiers.
There are many types of scopes, all can "see" both substances. So the answer depends upon the discipline your focus is in, if any.
An economical light microscope would allow a lay person to detect one from the other. A lot of valuable data there for sure.
Magnifications are limited to less than 1000 x but sufficient to discern either.

Scanning electron microscopy offers terrific 3d imagery and structural indices not detectable with other systems.
Transmission too, but those are expensive even as university turn overs.
enjoy.