First post, first image...

[shawngibson]

Hi all,

Been lurking for a few days, finally got my prepared slides, so I spent the evening working a bee leg. Never done this before, microscopes (and biology) are very new to me.

No official science background, but I've been reading science since I was a kid (I'm 46 now).

Glad I found this place...

Here's my first image, a bee leg. My own thoughts, somewhere there is a lack of resolution. There are no crisp edges. Gear is a 40x-2000x OMAX compound and 14mp OMAX USB 2 camera.

I have a photography background, and so my initial thoughts are that I need to replace the objectives AND the camera. This could get very expensive very quickly. I could use some advice, please and thank you. Anyways, here's the blob:



Thanks for letting me be here:)
Shawn

[einman]

Could be any of a number of issues.

Is this a trinoc scope?

Was the camera parfocal with the eyepieces?

Just a few questions ..pretty soon everyone will be jumping in..LOL

The lighting does not look right either. What model scope are you using?

[shawngibson]

Hi, and thanks:)

Not trinocular, it's binocular:

http://www.amazon.ca/gp/product/B005LIG ... ge_o00_s00

Was the camera parfocal with the lenses? I don't know. I know there was some sort of slide included with the 14mp camera, but OMAX didn't include any instructions.

Lighting is the microscope lighting with a pair of goosenecks on the top because the specimen wasn't transparent.

Shawn

[einman]

Bad news- Those cameras are not very good. But even so you should get a decent picture from it. Did you use the software on your computer to focus the eyepiece tube containing the camera before the photo?

What does the leg look like without the goosenecks? Trying to get a feel for the quality of the base illumination. Depending on how the slide was prepared the leg can be more transparent than you think.

I would use one or the other illumination initially to get a feel for the scope and camera etc.

[einman]

Given you are into photography I would buy an eyepiece adapter and use your current DSLR. It will most likely interface with the software as well.

[shawngibson]

>> Did you use the software on your computer to focus the eyepiece tube containing the camera before the photo?

No. Let's go one step at a time. How do I do this, please?

[shawngibson]

Given you are into photography I would buy an eyepiece adapter and use your current DSLR. It will most likely interface with the software as well.

I don't actually have a real camera right now. Too many hobbies:( The sensor on this OMAX unit looks like a point-and-shoot sensor. I'd love to put a Fuji X-E1 sensor onto the setup, great organic feel to it. Is that possible?

Shawn

[einman]

Hmmm.. okay load the software onto your laptop or desktop.

Once you have done that if not already boot up the program. You should see the image of the specimen on your screen.

Once you have an image you should look through the eyepiece w/o the camera and focus the image.

Then go to the software and focus the image on the screen using the eyepiece with the camera. Do not use the focus knobs.

That will make the camera and eyepiece parfocal that is both will be focused at the same time.

Take the pic..

[einman]

You may have to reverse the process by starting with the camera then finishing with the eyepiece if that makes sense. If you cant get the camera in focus.

[shawngibson]

Thanks Einman. I have literally an hour of experience with a microscope right now, so I'll read your words slowly lol.

My background in photography is with people. I've always kept it simple:

http://photo.net/photodb/member-photos?user_id=26697

...so this is a whole new world to me.

[einman]

Sorry-- Not used to walking someone through the process online.

[mrsonchus]

My little piece of advice is not to panic, definitely don't start buying more (replacement) kit! Most systems when optimized will give remarkably good results, and you're in the right place to get all the help and advice from this forum's many highly skilled and experienced members.
You can't expect to be good at this before you've spent time learning the techniques, pure application of technology won't do the job, just cost money and discourage - I know this from experience myself.
Keep it simple, start with a less problematic specimen, e.g. something that's transparent such as a plant-section - this will dramatically improve your chances of achieving a good image to start with, and this in turn will encourage and inspire you rather than discourage and frustrate you, I know, I've been through those stages and a tiny little good result can be a real encouragement!

My choice would be, for now, forget top-lighting, use only truly transparent and thin specimen with 'bottom lighting' to get you going. Practice with your 'scope to get a good image with the various objectives (lenses) before you even think about the added learning-curve of attaching a camera....

Good luck, use the experts of this forum, they're a very helpful and friendly bunch indeed!

[billbillt]

Actually, your photo is not THAT bad to me... I do know that the Omax stands can produce very fine and sharp images... Just keep experimenting would be my first suggestion..

BillT

[apochronaut]

For what you are photographing and the way you are lighting it, that isn't a terrible image. Microscope objectives are optimized to work with samples of specific thicknesses . The lower the magnification and N.A., the less critical this is but it is still there. LIghting from the top and below, can induce flare and cause a loss of contrast.

If you are unhappy with this image because you are comparing it to some images you are seeing here and on-line , you have to remember that a great number of photomicrographs these days are cleaned up in post processing, so such optical artifacts such as diffraction, flare and chroma have been scrubbed.

[shawngibson]

Thanks guys.

Yeah, I suppose I'm trying to run before I walk. The fact that I haven't calibrated anything yet, or even learned how to use the software, just call me Newb:)

I bought both a compound and a stereo/dissection microscope (both OMAX, and the stereo is trinocular). The dual gooseneck light is actually meant for the stereo scope. I have a dark field condenser coming for the compound.

I've purchased a bunch of books that should arrive end of October (Exploring With the Microscope: A Book of Discovery & Learning, Envisioning Science: The Design and Craft of the Science Image, and Practical Digital Photomicrography: Photography Through the Microscope for the Life Sciences).

I have about 20 arthropods coming from BioQuipBugs, and a couple dozen prepared slides, also mostly arthropods, mostly sections, and a dissection kit (a mollusc, a fish, a crayfish, a worm) coming as well.

A bunch of stains as well, and a kit with blank slides and some Canada Balsam for making my own slides.

Basically, I'm in this 100% but am starting with pretty much no knowledge.

I guess calibrating the camera/scope setup should be step one. Trying to figure out how to prep an ant for use with a microtome...maybe not so much right now lol.

Shawn

[gekko]

I cannot add anything useful to the excellent comments you got above other than say welcome to the forum.

[Dale]

Trying to figure out how to prep an ant for use with a microtome. ?? Aw, yer jokin right??

[shawngibson]

Yes:)

[shawngibson]

Second image. Followed advice above, largely. Single light source from the scope, with a Tiffen filter loosely placed over the condenser. 44 images focus stacked, software is Helicon Focus. No other adjustments.

Better, I hope...

Shawn

[shawngibson]

Holy Cow that's huge...sorry:(

[mrsonchus]

How about that! A good start! Just keep practicing, keep everything simple and concentrate on the very basic aspects of technique with both 'scope and camera - that's a good start and a pretty nicely executed picture, but I'd use a subject that you can recognize and reproduce if I were you, this would greatly aid your assessment of results and allow you to follow your early progress.

Perhaps something nice and flat, transparent and familiar, say a fly's wing for an animal (insect) subject or maybe some pollen from a plant. Either may be put into 50/50 water and (just plain old cake-making) glycerin, put onto a slide with a cover-slip (great practice) and viewed/photographed, also this glycerin/water mix will keep your specimen from drying-out or decaying for at least a week at room-temp, double that if put into a refrigerator - a 'standard-specimen' is definitely the way to start out.

You'll also find a significant enhancement to image-quality given by the glycerin/water mix's relatively high refractive index.

Good luck and congratulations on a good start - you're in for a great and hugely enjoyable adventure - the good folk of this forum will advise you about just about any aspect that you would like to master concerning microscopy - there's a considerable amount of expertise in this forum for you to learn alongside!

Keep up the good work and have fun!

[shawngibson]

Thank you so much, John B. I haven't been this excited, dedicated, and crazy-obsessed since I discovered girls when I was in high school lol!

[einman]

Girls? What are those?...hmm... let me check the index of my Ward's and Whipple..

[shawngibson]

You'll find them under "Crazy" in the dictionary. Just kidding...

[gekko]

To my eyes, your second image is quite good. My only suggestion for the future would be to place any filters below the condenser in order to allow the condenser to be brought up all the way (where it should generally be); if you need to reduce the aperture (for example, to increase contrast), use the condenser iris diaphragm rather than lower the condenser. This would be much more important with higher power (higher NA) objectives.

[shawngibson]

Thanks Gekko. Per your advice, filter in the correct spot. A dead midge, 72 images focus-stacked. Filter was Tiffen gel, yellow-orange-ish.

[billbillt]

WOW!!... That is very good!..

BillT

[JimT]

Shawngibson, welcome to the forum. Huge improvement in images between your first post and last one. Well done and keep 'em coming.

JimT

[Dale]

Wow. 72 images. Never thought about using my photo filters, still trying to see if I
can train an ant to use a microtome