40x doesn't show a clear image.

[WonderfulWafflez]

I just started learning about my microscope and trying to use it. I've figured it out mostly but I just can't use my 40x zoom or 100x zoom! I thought it was a focusing issue yet any troubleshooting tips I've tried hasn't changed the image. When I look through the 40x zoom it is just a gray image and I can see some microbes on the lens (I think) but the sample doesn't show up! When I switch it to 10x or 4x I can see the sample fine...

https://ibb.co/mcXh3Hq

The image is a little unclear since I took it with my phone...
More info on my setup:
I was using slides on my samples, .12 - .16 mm
I did try focusing the 40x really close to the sample
I tried to adjust the lighting
Using a compound microscope

Edit: originally I thought it was a focusing issue but now I'm not sure what it is...

[apochronaut]

The term " zoom" complicates things. I don't know what you are zooming or how. Zoom was invented by a mfg. of professional microscopes using a balanced but complex rearranegement of optics but became compromised with cheap optical systems that provided zoom at the expense of quality.
Nevertheless you should get an image of some kind.
You need to detail A) what you are looking at. B) what the system you are looking at it through is. C) What you have been smoking leading up to your post.

[WonderfulWafflez]

I'm sorry I meant 40x objective. Where I would look through the eyepiece to see the image I sent in the post.

And that smoking thing is a bit rude. I'm a beginner at using microscopes and I want help with my question.

[Alexander]

Some information about the microscope, the objectives and the samples would help to answer your question.

Do you use cover slides? How thick they are?

How are your samples embedded?

Do you use immersion oil? The 100x is most probably an immersion design.

The working distance of 40x and 100x objective is very tiny. Placing an embedded sample upside-down or even using a cover slide too thick would prevent one from focusing on the sample. Looking at the sample without a cover slide would give a quit unsharp picture but yo would see the sample. A 100x without immersion oil would give no picture. Same for a 40x spoiled with immersion oil.

There are more possible explanation. More information on the setup is required.

[GerryR]

Some information about the microscope, the objectives and the samples would help to answer your question.

Do you use cover slides? How thick they are?

How are your samples embedded?

Do you use immersion oil? The 100x is most probably an immersion design.

The working distance of 40x and 100x objective is very tiny. Placing an embedded sample upside-down or even using a cover slide too thick would prevent one from focusing on the sample. Looking at the sample without a cover slide would give a quit unsharp picture but yo would see the sample. A 100x without immersion oil would give no picture. Same for a 40x spoiled with immersion oil.

There are more possible explanation. More information on the setup is required.

Curious as to why you would expect no picture without immersion oil on 100x?? (My scopes both give a good image with no oil at 100x.)

[Alexander]

I own half a dozen 100x immersion objectives. Non of them produces anything that would qualify as a "good picture" while not immersed.

[apochronaut]

Using a liquid immersion with a refractive index greater than that of air, will increase the working distance of the objective by a factor equal to the positive ratio of the two n's. Depending on the thickness of the sample/coverslip and the original w.d. of the objective in it's specified immersion medium, using an immersion objective in air could cause the objective to hit the coverslip before it can focus, thus yielding no discernible image. This is more likely with an oil objective than water or glycerin.

Under conditions where an objective has a normal to longish working distance and the sample/coverslip parameters are within specification, oil immersion objectives will normally focus in air but since they rely on the immersion medium to gather more light due to an increase in the N.A. , plus the Increased N.A. results in maximum resolution the image will be darker than it should be and have low resolution. The effective N.A. of the objective will be below 1.
Objectives are also corrected for spherical aberration matching the type of immersion medium and w.d. Departing from the specified medium and thus altering the w.d. will result in an increase in spherical aberration. Triple whamy going against image quality.

Sometimes I will flip in a 100X without oil to get an interpretation of the details I am looking for but if I decide on actually using the objective for observation, I always oil the slide and often the çondenser too. It doesn't make sense to have a 1.3 or even a 1.0 oil objective for that matter and not use it's excellence. I have a 40X 1.0 oil planapo and in oil, it is an outstanding objective. In air: blah.

[WonderfulWafflez]

Some information about the microscope, the objectives and the samples would help to answer your question.

Do you use cover slides? How thick they are?

How are your samples embedded?

Do you use immersion oil? The 100x is most probably an immersion design.

The working distance of 40x and 100x objective is very tiny. Placing an embedded sample upside-down or even using a cover slide too thick would prevent one from focusing on the sample. Looking at the sample without a cover slide would give a quit unsharp picture but yo would see the sample. A 100x without immersion oil would give no picture. Same for a 40x spoiled with immersion oil.

There are more possible explanation. More information on the setup is required.

Yes I used cover slides, they are .12 mm thick.

[WonderfulWafflez]

Using a liquid immersion with a refractive index greater than that of air, will increase the working distance of the objective by a factor equal to the positive ratio of the two n's. Depending on the thickness of the sample/coverslip and the original w.d. of the objective in it's specified immersion medium, using an immersion objective in air could cause the objective to hit the coverslip before it can focus, thus yielding no discernible image. This is more likely with an oil objective than water or glycerin.

Under conditions where an objective has a normal to longish working distance and the sample/coverslip parameters are within specification, oil immersion objectives will normally focus in air but since they rely on the immersion medium to gather more light due to an increase in the N.A. , plus the Increased N.A. results in maximum resolution the image will be darker than it should be and have low resolution. The effective N.A. of the objective will be below 1.
Objectives are also corrected for spherical aberration matching the type of immersion medium and w.d. Departing from the specified medium and thus altering the w.d. will result in an increase in spherical aberration. Triple whamy going against image quality.

Sometimes I will flip in a 100X without oil to get an interpretation of the details I am looking for but if I decide on actually using the objective for observation, I always oil the slide and often the çondenser too. It doesn't make sense to have a 1.3 or even a 1.0 oil objective for that matter and not use it's excellence. I have a 40X 1.0 oil planapo and in oil, it is an outstanding objective. In air: blah.

But would this be the explanation why I can't see a clear image with the 40x objective?

[WonderfulWafflez]

I own half a dozen 100x immersion objectives. Non of them produces anything that would qualify as a "good picture" while not immersed.

Yes but 40x objectives shows the same pictures as the 100x, but I need to test the 100x with immersion oil...

[Alexander]

Yes but 40x objectives shows the same pictures as the 100x, but I need to test the 100x with immersion oil...

To answer your question more information about the equipment used is needed.

[WonderfulWafflez]

Yes but 40x objectives shows the same pictures as the 100x, but I need to test the 100x with immersion oil...

To answer your question more information about the equipment used is needed.

Well I'm using a compound microscope, I should have said that in the post sorry.

[Alexander]

Well I'm using a compound microscope, I should have said that in the post sorry.

What type and make?
Exact description of all objectives?
What type of condenser? How is it handled?
Illumination?
Procedures? Procedures? Procedures?

This is my last try. If not successful, I will give up.

[apochronaut]

Using a liquid immersion with a refractive index greater than that of air, will increase the working distance of the objective by a factor equal to the positive ratio of the two n's. Depending on the thickness of the sample/coverslip and the original w.d. of the objective in it's specified immersion medium, using an immersion objective in air could cause the objective to hit the coverslip before it can focus, thus yielding no discernible image. This is more likely with an oil objective than water or glycerin.

Under conditions where an objective has a normal to longish working distance and the sample/coverslip parameters are within specification, oil immersion objectives will normally focus in air but since they rely on the immersion medium to gather more light due to an increase in the N.A. , plus the Increased N.A. results in maximum resolution the image will be darker than it should be and have low resolution. The effective N.A. of the objective will be below 1.
Objectives are also corrected for spherical aberration matching the type of immersion medium and w.d. Departing from the specified medium and thus altering the w.d. will result in an increase in spherical aberration. Triple whamy going against image quality.

Sometimes I will flip in a 100X without oil to get an interpretation of the details I am looking for but if I decide on actually using the objective for observation, I always oil the slide and often the çondenser too. It doesn't make sense to have a 1.3 or even a 1.0 oil objective for that matter and not use it's excellence. I have a 40X 1.0 oil planapo and in oil, it is an outstanding objective. In air: blah.

But would this be the explanation why I can't see a clear image with the 40x objective?

Not likely. Oil wouldn't enter into the picture for most 40X objectives. I just used the above as an illustration of how important oil is to an oil immersion objective. I assume yours is a dry objective and likely .65 N.A.?
If so, you should be able to distinctly see two cocci as separate when they are touching , since a correctly working .65 objective under good conditions of slide preparation should resolve to about 500 nm or .5 micron.

A good way to test that is to selectively breed some bacteria you know the size of. Take a small plastic çontainer with an airtight lid and wash it well. Put 1/2 c. water in it and 1 level tsp. of salt. You now have an approx. 5% salt brine. Your container should be small enough to provide about 3/4 to 1" of brine. Double the recipe if you need to but it is best to have a container deeper than wide. One of those 250gm. or 1/2 lb. cream/cottage cheese type containers is good but make sure it is immaculately washed. Add a 2 or 3 sliced up radishes or a sliced up baby cucumber or a piece of zuchinni sliced up or 4 or 5 1/4" slices of broccoli stem, or a hand sized chunk of cabbage leaf sliced up. Some firm vegetable that hasn't been scrubbed to death, just rinsed off.
Put some flat object that will fit inside the i.d. of your container on them to keep the vegetables under the surface: a small plate, inverted plastic lid, take out beverage lid inverted, can lid, glass disc ,thin scrubbed flat stone if you can find one. If the object is too light, put some small scrubbed stones or some washed alleys on it. Anything relatively inert.
The whole effort should take no more than 15 minutes., once you assemble the few bits of hardware.
Keep at 80° F. +- 5 or so for about 3-4 days. You will likely see a bit of cloudiness in the brine by then. The first bacteria to grow will be Leuconostoc Mesenteroides and that is what you will have It will be in small chains of a few to maybe 20. Each individual is approx. .6 microns across, slightly oval up to 1.2 microns long. Your .65 objective should easily distinguish them as separate.
Put the vegetables in a salad or on a sandwich unless you used a lead disc to weigh them down. They are good for you.

[WonderfulWafflez]

But could this be a main issue with why I can't see anything whatsoever...

[deBult]

People are trying to help, but evidently ….

[apochronaut]

But could this be a main issue with why I can't see anything whatsoever...

The post implied that you aren't getting a clear image but you are saying actually, no image is the problem.
Can you explain the reference to "zoom". Normally a zoom feature is in the optical section above the objective and is used with each objective in turn but you mention 10X zoom and 40X zoom. Can you explain what you mean by that? Maybe post a picture or two.

If it is truly a zoom objective of some kind, I would include some gross mechanical fault in the mechanism as a possibility. A displaced optic for instance. If it is just a standard objective, then that is unlikely.
You haven't mentioned whether this is a new or used microscope. The only reason I mention that is that some used microscopes are on the market because they are defective in some way.
I assume you have ruled out a heavy layer of gunk or residue on the front lens?

[BramHuntingNematodes]

Hmm Zeiss had a low power zoom objective. B&L had the student zoom scope that went up to 40x I think, and of course the low power flip-bottom lens.

I would think it's a parfocal issue, and the user doesn't quite have enough experience to focus the 40x. The range over which a 40x will focus on a thin sample is very small. You might try lower the lens until it is touching the coverslip while looking sideways at the stage, then start looking through the oculars as you very gradually rack the lens up. Try it a few times, if there is still nothing your lens may be at fault. You can unscrew it and look through it pointed at a light fixture to see if there is a gross amount of damage.

[SWmicro]

I can see the sample fine...
https://ibb.co/mcXh3Hq
The image is a little unclear since I took it with my phone...

Firstly
I would not play with the 100x objective until we can sort out the 40x (and/or any other problems !), the 100x is very tricky to use at the best of times and you may well cause more damage than good because it must be placed very close to the slide/coverslip.

At first I thought you had posted a picture of nothing, only a white circle, But after a big stretch in Gimp some smudges became evident. Some are, I think, artifacts of the glass, others may be diagnostic like :-
- do any of the smudges move ?
a) when you rotate the eyepiece(ocular),
or
b) as you move the slide back and forward / side to side.

If b) , then do they change shape as you move the focus up and down ?

1.jpg (106.24 KiB) Viewed 5105 times

[LouiseScot]

It might be an idea to try and focus on just a thin piece of tissue in place of a slide. That would prevent any damage should the objective make contact. The working distance of a 40x objective is typically around 0.6mm.

Louise

[SWmicro]

Perhaps a high contrast item, like a thread, on the stage in place of the slide to see if you can see it moving and get idea of the 40x focus point.

[apochronaut]

He says he is using .12 coverslips. I don't know of any 40X that won't focus with those.

[SWmicro]

He says he is using .12 coverslips. I don't know of any 40X that won't focus with those.

That assumes the 40x even has the ability to focus. It may be too badly damaged already.

We just need more info from W W to help plot a path of discovery !

[WonderfulWafflez]

But could this be a main issue with why I can't see anything whatsoever...

The post implied that you aren't getting a clear image but you are saying actually, no image is the problem.
Can you explain the reference to "zoom". Normally a zoom feature is in the optical section above the objective and is used with each objective in turn but you mention 10X zoom and 40X zoom. Can you explain what you mean by that? Maybe post a picture or two.

If it is truly a zoom objective of some kind, I would include some gross mechanical fault in the mechanism as a possibility. A displaced optic for instance. If it is just a standard objective, then that is unlikely.
You haven't mentioned whether this is a new or used microscope. The only reason I mention that is that some used microscopes are on the market because they are defective in some way.
I assume you have ruled out a heavy layer of gunk or residue on the front lens?

I did post a picture. Click the link in the og post. I got this microscope about a year ago. It was a new microscope then.

[WonderfulWafflez]

I can see the sample fine...
https://ibb.co/mcXh3Hq
The image is a little unclear since I took it with my phone...

Firstly
I would not play with the 100x objective until we can sort out the 40x (and/or any other problems !), the 100x is very tricky to use at the best of times and you may well cause more damage than good because it must be placed very close to the slide/coverslip.

At first I thought you had posted a picture of nothing, only a white circle, But after a big stretch in Gimp some smudges became evident. Some are, I think, artifacts of the glass, others may be diagnostic like :-
- do any of the smudges move ?
a) when you rotate the eyepiece(ocular),
or
b) as you move the slide back and forward / side to side.

If b) , then do they change shape as you move the focus up and down ?

1.jpg

No they don't move, so I think it's just stuff on the objective lens.

And sorry for being difficult, I'm new at all of this.

[apochronaut]

Could you post a picture of the objective and the objective mounted on the microscope. Did the objective ever work properly, since you purchased it?

[WonderfulWafflez]

(I don't think the image button is working)

When I first got it the microscope I didn't use until when I started using it a couple weeks ago. So the objective never really has been working.

Could the objective be damaged?

[Alexander]

When I first got it the microscope I didn't use until when I started using it a couple weeks ago. So the objective never really has been working.

Could the objective be damaged?

Most probably not. A user problem is much more likely.

A possible source of your difficulties is the handling of the microscope's condenser. Does it have a front-lens that could be slipped away or some other lens that could be turned in and out?

Again:
To identify your problem and to provide a solution to it, a proper description of your working process is needed.

[WonderfulWafflez]

Yes there is a condenser and I've already tried adjusting it.

This whatci do when I use my microscope

-take the dust cover off
-turn on the microscope light
-carefully put in a slide and make sure it is latched in
-line up the sample

That's pretty much it...

[apochronaut]

The condenser will not stop an objective from focusing. It will only affect the intensity of illumination and the degree of diffraction and other aberrations and distortions that alter the illumination of the subject. The objective will still focus an image , although it might be a crappy image.