First water bear found elsewhere

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Lilly Begonia
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First water bear found elsewhere

#1 Post by Lilly Begonia » Sun Mar 04, 2018 11:08 am

I found this in tree lichens I got from Rotary Park here in the cape. This is the first water bear I've found that was not from the lichen on the tree behind my house, so it seems wherever I find lichens I can find water bears in them. I tried to get a stacked image of him, but then he started moving around. I enlarged these shots as Michael suggested but I think it blurs them a bit and the sharp focus is somewhat lost.

Image

Image

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mrsonchus
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Re: First water bear found elsewhere

#2 Post by mrsonchus » Sun Mar 04, 2018 12:36 pm

Wow, Lily these are super images, really detailed and most interesting to see! :D

You're building a good collection of images and knowledge my friend, keep them coming, they're great to follow. Your image-quality is very good as far as I'm concerned - it's really not easy to image such live specimens - let-alone moving ones!

Great adventures, please keep them coming.

John B. :)
John B

Lilly Begonia
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Re: First water bear found elsewhere

#3 Post by Lilly Begonia » Sun Mar 04, 2018 12:42 pm

mrsonchus wrote:Wow, Lily these are super images, really detailed and most interesting to see! :D

You're building a good collection of images and knowledge my friend, keep them coming, they're great to follow. Your image-quality is very good as far as I'm concerned - it's really not easy to image such live specimens - let-alone moving ones!

Great adventures, please keep them coming.

John B. :)
Thank you so much John! I just found the second one in that lichen collected yesterday, and here it is:

Image

I'm pretty happy with the detail in this one, and I really love that stacking software. This sure beats sitting here listening to my hair grow :)
Image

...and here's another one, third so far today...

Image

MichaelG.
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Re: First water bear found elsewhere

#4 Post by MichaelG. » Sun Mar 04, 2018 2:00 pm

Lilly Begonia wrote:I enlarged these shots as Michael suggested but I think it blurs them a bit and the sharp focus is somewhat lost.
Could I just check, Lilly ?
... How are you enlarging the images ?

When I suggested getting the bear bigger in the frame, I was thinking of higher magnification in the microscope; and you would of course then get less depth of field, and need to stack more shots.

It's all a rather delicate balance, especially when the posted images are of limited resolution.

MichaelG.
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Lilly Begonia
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Re: First water bear found elsewhere

#5 Post by Lilly Begonia » Sun Mar 04, 2018 2:09 pm

MichaelG. wrote:
Lilly Begonia wrote:I enlarged these shots as Michael suggested but I think it blurs them a bit and the sharp focus is somewhat lost.
Could I just check, Lilly ?
... How are you enlarging the images ?

When I suggested getting the bear bigger in the frame, I was thinking of higher magnification in the microscope; and you would of course then get less depth of field, and need to stack more shots.

It's all a rather delicate balance, especially when the posted images are of limited resolution.

MichaelG.
Well Michael, I have a 100x objective a 400x objective for this pursuit, and nothing in between those. So I resorted to using Preview on my Mac to enlarge the images. Clearly a 200x objective would have yielded better results, but I don't have one. Often the 100x objective gives too small an image, and the 400x cannot get the whole animal in the frame, so yes, it's a delicate balance, and bear in mind I've been at this about a month now.

How's this for resolution:

Image
Image

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Re: First water bear found elsewhere

#6 Post by MichaelG. » Sun Mar 04, 2018 4:20 pm

Couldn't ask for much better than that, Lilly

MichaelG.
.
One point that I must mention:
Unless I am very much mistaken, the two objectives that you mention are actually 10x and 40x
Omax specifies a range of magnifications, but these numbers are the product of 'objective x eyepiece'
http://www.microscopenet.com/omax-40x20 ... -9100.html

When you are using the USB 'eyepiece camera' the microscope eyepiece is not there.
It would be worth checking [*] what size your camera is seeing.

[*] Edit: I see that the camera comes with a 0.5x reduction lens and a 'calibration slide'
http://omaxmicroscope.com/microscope-ac ... linux.html
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Lilly Begonia
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Re: First water bear found elsewhere

#7 Post by Lilly Begonia » Sun Mar 04, 2018 11:00 pm

MichaelG. wrote:Couldn't ask for much better than that, Lilly

MichaelG.
.
One point that I must mention:
Unless I am very much mistaken, the two objectives that you mention are actually 10x and 40x
Omax specifies a range of magnifications, but these numbers are the product of 'objective x eyepiece'
http://www.microscopenet.com/omax-40x20 ... -9100.html

When you are using the USB 'eyepiece camera' the microscope eyepiece is not there.
It would be worth checking [*] what size your camera is seeing.

[*] Edit: I see that the camera comes with a 0.5x reduction lens and a 'calibration slide'
http://omaxmicroscope.com/microscope-ac ... linux.html
Okay, well being so new to this as I am I will take your word for it, and in fact that does make sense. I do have that calibration slide but I've been so busy working 22 hour shifts, dealing with maintenance (life is 90% maintenance) and chasing water bears I've not even looked into what it was or how to use it. It was just included, with no mention of what it was.

My microscope was billed as having 2000x magnification with an extra set of eyepieces that were included, but early on I learned that these were just empty magnification that only made the image larger with no additional detail, so those went into a box in the closet. Additionally my scope has been having focusing issues, and the software my camera came with, ToupLite for Mac is described by the manufacturer as "bare bones" after I wrote them about it's problems. It cannot do any sort of acceptable video and has nowhere near the features of ToupView for Windows. So I find myself laboring under certain handicaps.

You seem to really know your stuff, and I just got into this 21 days ago, so I do appreciate any advice you care to offer. I'd really like to do this right. How do I use that calibration slide? No instructions came with it, and no documentation came with ToupLite either. Bare bones indeed.

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mrsonchus
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Re: First water bear found elsewhere

#8 Post by mrsonchus » Sun Mar 04, 2018 11:12 pm

Hi Lily, calibration for measurement using Toupview with a Toupcam is pretty straightforward and one calibration needs to be made (only
needs to be done once, not every time) for each objective...
Last year I posted the method in this forum, here's a link to my post re calibration for measurements with Touview...

Hope it helps.

John B. :)
John B

Lilly Begonia
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Re: First water bear found elsewhere

#9 Post by Lilly Begonia » Sun Mar 04, 2018 11:26 pm

mrsonchus wrote:Hi Lily, calibration for measurement using Toupview with a Toupcam is pretty straightforward and one calibration needs to be made (only
needs to be done once, not every time) for each objective...
Last year I posted the method in this forum, here's a link to my post re calibration for measurements with Touview...

Hope it helps.

John B. :)
I don't have ToupView, that's for Windows, I have ToupLite for Mac, and I really do not trust it. It's been a real bad experience in more than one way. I am heavily invested in the Mac platform, and it's looking like to get any decent microscope camera output I'll be forced to buy a windows box, on top of the expense of my microscope and camera. Not happy about that.

The manufacturer of my microscope tells me:

As for feature availability, the development team has already added many of the Windows features into the Mac software. Initially (a couple years ago), the application was very barebones, and only had the capability for basic live preview and image capture. Over time, they've been adding features (such as video recording and more improved controls). It is in my best educated guess that they'll eventually roll out additional features into the Mac software, given their history of improving the software. As for a timeline for particular features, I do not have that information.

Yeah, right, video recording, and of only the very worst quality that is flat-out unusable. They will eventually roll out improvements... sure they will. It's Chinese software, and they appear not to care at all. I'm not satisfied with EVENTUALLY!

MichaelG.
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Re: First water bear found elsewhere

#10 Post by MichaelG. » Mon Mar 05, 2018 7:09 am

Lilly,

I must first emphasise [shout from the roof-tops] that I think you are doing a fantastic job, despite the obvious frustration with your chosen equipment.

To understand the calibration, it's probably worth starting with the camera sensor:
This is specified as having 2560x1922 pixels, on a pitch of 2.2 microns
[ micron = micrometre = 0.001 mm ]
... So its active area is roughly 5.6 mm x 4.2 mm

Width : 2560 x 2.2 = 5632 microns
Height : 1922 x 2.2 = 4228.4 microns

The magnification of a nominal 10x objective may not be exactly that, so its worth using the calibration slide ... Just put it on the stage and check what the width of the field is; both visually, and with the camera [with and without its 0.5x 'reduction lens'].

You will, I am very sure, quickly get to grips with this; but if you need any further explanation, we're all here to help.

Obviously; pictures that are printed, or viewed on screen, have their own magnification applied to the physical size of the camera sensor !! ... This is why it's best practice to include a scale-bar in published images.

Keep up the great work ... Your enthusiasm and your results are inspirational.

MichaelG.
Too many 'projects'

Lilly Begonia
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Re: First water bear found elsewhere

#11 Post by Lilly Begonia » Mon Mar 05, 2018 11:37 am

MichaelG. wrote:Lilly,

I must first emphasise [shout from the roof-tops] that I think you are doing a fantastic job, despite the obvious frustration with your chosen equipment.

To understand the calibration, it's probably worth starting with the camera sensor:
This is specified as having 2560x1922 pixels, on a pitch of 2.2 microns
[ micron = micrometre = 0.001 mm ]
... So its active area is roughly 5.6 mm x 4.2 mm

Width : 2560 x 2.2 = 5632 microns
Height : 1922 x 2.2 = 4228.4 microns

The magnification of a nominal 10x objective may not be exactly that, so its worth using the calibration slide ... Just put it on the stage and check what the width of the field is; both visually, and with the camera [with and without its 0.5x 'reduction lens'].

You will, I am very sure, quickly get to grips with this; but if you need any further explanation, we're all here to help.

Obviously; pictures that are printed, or viewed on screen, have their own magnification applied to the physical size of the camera sensor !! ... This is why it's best practice to include a scale-bar in published images.

Keep up the great work ... Your enthusiasm and your results are inspirational.

MichaelG.
Thank you so very much Michael! Well I got out that calibration slide, put it on the stage, turned to my lowest power objective and focused on it. Here's what I see in the middle of the circle:

Image

There are no numbers there. I'm not sure how to use it. I'm a bit over my head with this, but I'm not giving up. I've been reading and searching around and I find mention of the stage micrometer and the eyepiece micrometer. I take the stage micrometer to mean the markings on the top and right of the stage, but I'm not sure what the eyepiece micrometer is, none appears in the eyepiece, unless it's that image on the slide. I looked at YouTube videos on how to calibrate a microscope, but they look very different from what I have and are for a variety of other scopes and calibration slides, so I'm a little lost at this point.

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Re: First water bear found elsewhere

#12 Post by MichaelG. » Mon Mar 05, 2018 1:49 pm

Lilly Begonia wrote:
Thank you so very much Michael! Well I got out that calibration slide, put it on the stage, turned to my lowest power objective and focused on it. Here's what I see in the middle of the circle:

Image

There are no numbers there. I'm not sure how to use it. I'm a bit over my head with this, but I'm not giving up. I've been reading and searching around and I find mention of the stage micrometer and the eyepiece micrometer. I take the stage micrometer to mean the markings on the top and right of the stage, but I'm not sure what the eyepiece micrometer is, none appears in the eyepiece, unless it's that image on the slide. I looked at YouTube videos on how to calibrate a microscope, but they look very different from what I have and are for a variety of other scopes and calibration slides, so I'm a little lost at this point.
Lilly,

Your 'calibration slide' is the 'stage micrometer'
The size of the minor divisions is stated to be 0.01 mm [see the microscope spec.]

... an 'eyepiece micrometer' is sometimes useful, but is a Red Herring in the present discussion.

Sorry, can't write more just now, but I'll be back this evening [U.K. time]

MichaelG.
Too many 'projects'

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Re: First water bear found elsewhere

#13 Post by MichaelG. » Mon Mar 05, 2018 5:50 pm

Hello again, Lilly

This is very rough, but should give you the general idea:
100 divisions of 0.01mm
100 divisions of 0.01mm
IMG_1847.JPG (403.1 KiB) Viewed 4729 times
... obviously it would be better to align the scale with the camera's horizontal.

MichaelG
Too many 'projects'

Lilly Begonia
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Re: First water bear found elsewhere

#14 Post by Lilly Begonia » Mon Mar 05, 2018 6:18 pm

MichaelG. wrote:Hello again, Lilly

This is very rough, but should give you the general idea:

... obviously it would be better to align the scale with the camera's horizontal.

MichaelG
Yes it would Michael, the camera slips around in the eyepiece tube, even after I zip-tied it's cable down. I wish there was a set screw in that tube.

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Re: First water bear found elsewhere

#15 Post by MichaelG. » Mon Mar 05, 2018 6:54 pm

Lilly Begonia wrote:
MichaelG. wrote: Yes it would Michael, the camera slips around in the eyepiece tube, even after I zip-tied it's cable down. I wish there was a set screw in that tube.
Here's one of life's little miracles >>> https://en.m.wikipedia.org/wiki/Blu_Tack

MichaelG.
Too many 'projects'

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Jonnyvine
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Re: First water bear found elsewhere

#16 Post by Jonnyvine » Wed Mar 14, 2018 4:19 pm

Cool! I've been trying to look for these myself. :)

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