Amoeba (video)

Here you can post pictures and videos to show others.
Post Reply
Message
Author
User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Amoeba (video)

#1 Post by gekko » Wed Oct 15, 2014 12:38 am

This is an old video; I post it because it is colorful (in contrast to the scuticocilliate video which was devoid of color). I think this might be Thecamoeba. Objective: 20x; illumination: DIC; camera: Olympus E-P1; duration: about 3 minutes (please click on vimeo to view in HD).


User avatar
75RR
Posts: 8207
Joined: Sun Oct 12, 2014 2:34 am
Location: Estepona, Spain

Re: Amoeba (video)

#2 Post by 75RR » Wed Oct 15, 2014 3:36 pm

Very nice. I find Amoebas fascinating.
It is also encouraging that you are making video of this high quality with your E-p1.
Can't get the White Balance sorted out in movie mode on my E-p2. Looking forward to getting it right.
If I end up making videos like yours I will be very happy.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: Amoeba (video)

#3 Post by gekko » Wed Oct 15, 2014 7:40 pm

Many thanks, 75RR.
On my E-P1, what I do is set "custom white balance" in still mode (I use Aperture Priority because I don't use an attached camera lens) using a blank (clear) area on the slide. I'm not 100% sure, but I've been assuming that the white balance set in still mode also applies to video. Also, I found out (I really need to read the manual!!!) that I can adjust exposure in video the same way as in still mode by adjusting the bias value before I start shooting the video. I do this by pressing the INFO button (I assume this is the same on the E-P2) repeatedly until the "live" histogram is displayed, and then adjustthe bias value to keep the right-most end of the histogram curve away from the right edge of the histogram window (to avoid overexposure).

User avatar
75RR
Posts: 8207
Joined: Sun Oct 12, 2014 2:34 am
Location: Estepona, Spain

Re: Amoeba (video)

#4 Post by 75RR » Wed Oct 15, 2014 8:04 pm

Thanks gekko, will "endeavor to persevere".
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

lilewis
Posts: 9
Joined: Wed Oct 22, 2014 11:51 pm

Re: Amoeba (video)

#5 Post by lilewis » Thu Oct 23, 2014 12:07 am

gekko, I am new to microscopy. I purchased an Amsco B120 with the 10mp camera, but also have a couple of DSLR's (Olympus E5 and OM-D E-M1) and an older Oly C5050Z I could use and I believe all would yield better resolution than the Amsco USB camera.

Being new to this, I was happy with the Amscope, but after viewing the images on this forum, I'm beginning to think I won't be able to photograph the level of detail I see that you and others here do.

I do have one question about a post you made above. You said you don't use the camera lens? Is that just for taking the white balance reading? I assume you use the lens for the shots/videos. Or is there something I don't know about proper setup with the microscope?

User avatar
Oliver
Posts: 187
Joined: Sat Oct 11, 2014 12:57 pm
Location: Austria
Contact:

Re: Amoeba (video)

#6 Post by Oliver » Thu Oct 23, 2014 10:00 am

I believe all would yield better resolution than the Amsco USB camera.
10MP is a lot, much more than you "need". Do not forget, that many microscope objectives and specimens are not even able to provide the quality that the camera is able to capture (in other words, not the camera is the problem in most cases, it is the quality of the specimen and set-up).

Oliver
Image Oliver Kim - http://www.microbehunter.com - Microscopes: Olympus CH40 - Olympus CH-A - Breukhoven BMS student microscope - Euromex stereo - uSCOPE MXII

lilewis
Posts: 9
Joined: Wed Oct 22, 2014 11:51 pm

Re: Amoeba (video)

#7 Post by lilewis » Thu Oct 23, 2014 10:48 am

Hi Oliver,
I originally bought Amscope's 1.3mp camera as was very unhappy with it (I should have known better in the first place), so I decided to just make the jump to the 10mp. But, correct me if I'm wrong, most of the really good photos I see on this and other groups seems to be taken by DSLRs, (and I supposed with higher quality microscopes).

I do have a stacked photo of a fruit fly that I think came out ok:
https://www.flickr.com/photos/lilewis/15506833356/
but many of my others didn't come out so well and I have not yet been able to capture amoebae(sp?) to the detail I see on other photos. But probably that's because, now that I think about it, I'm still only comfortable using the 4x and 10x objectives.

I did learn something last night from The QCC on your site that I think may be helpful regarding how to use the condenser aperture.

I'm a newbie to microscopes and know that I have a lot to learn.

User avatar
Oliver
Posts: 187
Joined: Sat Oct 11, 2014 12:57 pm
Location: Austria
Contact:

Re: Amoeba (video)

#8 Post by Oliver » Thu Oct 23, 2014 11:26 am

But, correct me if I'm wrong, most of the really good photos I see on this and other groups seems to be taken by DSLRs, (and I supposed with higher quality microscopes).
DSLRs do indeed have one significant advantage over dedicated microscope cameras and this is that they generally have a much higher dynamic range. This means that bright parts of the specimen do not appear to be washed out and totally white, and dark parts are not completely black. I think it is not always a resolution issue. One disadvantage of DSLRs is that you need specific adapters and optics, which can handle the larger sensor size (much easier to get un-clear corners due to larger sensor). There is also the problem of vibration. The advantage is that you can do HD videos with many DSLRs (USB2 microscope cameras are too slow).

Köhler illumination sometimes improves quality, especially with higher magnification, by reducing stray light and thereby increasing contrast. Condensor diaphragm is super important in controlling contrast and depth of field. And contrast enhancement and background dust removal really work miracles in improving the subjective impression of the image (even if the resolution is not improved)

Try the following: Take a permanent mount (no movement) and take a picture using your 10MP and 1.3MP camera. Then enlarge the 1.3MP image to a size of 10 MP and view both 10MP images (the real one and the enlarged one) at normal resolution (!) side by side. You can then see if the 10MP camera was able to capture details that the 1.3 MP camera was not. This allows you to assess to what extent resolution plays a role. With 1.3MP it will certainly play a role.

You can also take overlapping images and then stitch them together (using MS ICE, free) to make a larger picture, which you can then again reduce in size. This sometimes also gives an impression of a better image, because by reducing the image size you also lose some distracting details and noise. But much depends on the actual specimen.

Then try: Take a picture of the same specimen using both cameras and reduce both images to eg 600x400 and compare them side by side. The images published in this forum are rarely larger. So you are essentially removing resolution for the Web.

You might be interested in the article I wrote in the first issue of the magazine ("The power of digital post processing"): http://www.microbehunter.com/microbehun ... uary-2011/

Oliver.
Image Oliver Kim - http://www.microbehunter.com - Microscopes: Olympus CH40 - Olympus CH-A - Breukhoven BMS student microscope - Euromex stereo - uSCOPE MXII

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: Amoeba (video)

#9 Post by gekko » Thu Oct 23, 2014 12:00 pm

First, may I suggest, if you have not done so already, that you look at the Microbe Hunter main website where Oliver has provided a lot of very helpful and authoritative information on the subject.

If I may try to answer some of your questions briefly: there are two methods commonly used to connect a camera to a microscope:
(1) The camera with its attached lens is used simply to replace the operator's eye, so-called afocal method, where you position the camera to look through the eyepiece. This may be the more convenient method with a binocular (as opposed to trinocular) microscope.
(2) The camera is used without a camera lens, and a projection lens in the microscope projects the light to form an image directly onto the camera's sensor (this is the method I use, mainly because I find it more convenient than the afocal method, and I have a trinocular microscope).
Both methods are capable of excellent results.
An excellent, authoritative article on connecting a camera to a microscope:
http://krebsmicro.com/photomic1/photomic1.html
Other very useful related articles as well as amazing photomicrographs:
http://www.krebsmicro.com/

I see no reason why your Amscope should not give you excellent images, but, as with everything, gaining some experience and getting some practice in the use of the microscope are important. However, there are a few points that come to mind that affect the quality of the microscope image:
(1) It is obvious that focusing the microscope on the object on the slide should be done very carefully.
(2) With higher power objectives, the thickness of the cover glass is important (using an inappropriate cover glass can result in poor images due to increased spherical aberration).
(3) Correct adjustment of the condenser and its aperture are critical. Briefly, for microscopes such as your Amscope 120B, the condenser should always be raised the top limit of its travel (assuming that you are using the condenser that was designed for your microscope). Then, with the microscope focused on the object, a quick method of setting the condenser iris is to open it all the way, then gradually close it while looking through the eyepieces until you see a very slight reduction in the light level. This should be a good starting point from which you can either close it a bit more or open it a bit to obtain the best image (best compromise between resolution and contrast). The adjustment of the iris opening should be done whenever a different objective is used and even, using the same objective, when a different object having different contrast is being observed.
(4) The camera should be located such that when the image, as seen by eye through the eyepiece, is in focus, the image on the camera's sensor is in focus at the same time. If the microscope focus points for visual work and photography are different, the quality of the captured image will suffer (due to an increase in spherical aberration).

I hope this helps, and good luck with your new, wonderful hobby!

lilewis
Posts: 9
Joined: Wed Oct 22, 2014 11:51 pm

Re: Amoeba (video)

#10 Post by lilewis » Thu Oct 23, 2014 3:24 pm

Oliver and Gekko,

Thanks for all the good information. I just read your magazine article and found it full of great advice.

I returned the 1.3mp USB camera to Amscope for credit towards the 10mp version so I can't do some of the tests you suggested. But I can and will compare some equal sized photos from each.

"the condenser should always be raised the top limit of its travel (assuming that you are using the condenser that was designed for your microscope)"
I didn't know that. Thx. The condenser came from Amscope with the microscope so I assume it's the correct one.

I'm encouraged now, and will set the condenser high and work the aperture as you both suggesed.

lilewis
Posts: 9
Joined: Wed Oct 22, 2014 11:51 pm

Re: Amoeba (video)

#11 Post by lilewis » Thu Oct 23, 2014 3:35 pm

Gekko,
I just finished the rest of your post and I very much appreciate the info both of you have given me.
I wish there were a local night school class on the stuff. There's a lot to learn to do it the right way.

I've opened the two Krebs articles and will have to read them later. Thanks again.

lilewis
Posts: 9
Joined: Wed Oct 22, 2014 11:51 pm

Re: Amoeba (video)

#12 Post by lilewis » Thu Oct 23, 2014 8:25 pm

I just read the 2 Charles Krebs links. All I can say is: WOW.
I now understand the concept of using a DSLR without a lens.

I'm interested in both 1 celled organisms, and insects. Although I was advised by Amscope that it would be preferable to look at insects with a stereo, I thought I'd try to work with the compound microscope that I just purchased, first.

I cut some ping pong balls in half and then cut a hole through the center of each half to insert the 'domes' over the objectives so I could diffuse a couple of gooseneck lamp led's onto the insects but the results are still harsh lighting and limited room to focus, in addition the the lamps using a lot of the table space.

I'm now wondering if I need one of each. Am I correct? And if so can I get a decent Stereo for $250 or less? I'm thinking Amscope again if for nothing else, commonality in eyepieces, etc. but am open to suggestions.

I'm not sure if I will pull the trigger on this - but I am interested in your comments.

Thank you

Loren

User avatar
Oliver
Posts: 187
Joined: Sat Oct 11, 2014 12:57 pm
Location: Austria
Contact:

Re: Amoeba (video)

#13 Post by Oliver » Thu Oct 23, 2014 8:39 pm

Insects with a compound microscope are possible, but only if they are sufficiently small and transparent. Otherwise you see only black. Wings of flies, bees etc. are great to look at.
To brighten up the insects, you can place them in a solution of KOH (others also use NaOH) or lactic acid. If they stay in there too long, then they will dissolve. Be careful when you use these substances.

Gooseneck lamps, LEDs: yes this works, especially with low magnification. The high mag objectives cast a shadow. A LED flashlight also works, but is far less convenient. Instead of pingpong balls you can also use a ring of paper around the specimen. Using 1 lamp will cast shadows and this can increase the impression of depth.
Oliver
Image Oliver Kim - http://www.microbehunter.com - Microscopes: Olympus CH40 - Olympus CH-A - Breukhoven BMS student microscope - Euromex stereo - uSCOPE MXII

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: Amoeba (video)

#14 Post by gekko » Thu Oct 23, 2014 11:48 pm

lilewis wrote:"the condenser should always be raised the top limit of its travel (assuming that you are using the condenser that was designed for your microscope)"
I didn't know that. Thx. The condenser came from Amscope with the microscope so I assume it's the correct one.
I would like to clarify that this is true for microscopes, like yours, that do not support Koehler illumination (for microscopes with Koehler, the condenser is adjusted as The QCC tried to explain in a recent post). In either case, the aim is to form a focused image of the condenser aperture at the back focal plane of the objective, such that the size of the condenser aperture limits the aperture of the objective to direct light. In other words, the plane of the condenser aperture and of the objective back focal plane are optical conjugates (see for example the link below):
http://www.microscopyu.com/articles/for ... ugate.html
I would like to add that, by the same argument, dark field stops, oblique illumination stops, Rheinberg filters, and the like are best located at (or as close as practicable) to the plane of the condenser diaphragm (in many condensers, a filter tray is provided immediately below the condenser iris diaphragm for the purpose).

lilewis
Posts: 9
Joined: Wed Oct 22, 2014 11:51 pm

Re: Amoeba (video)

#15 Post by lilewis » Fri Oct 24, 2014 12:47 am

Oliver, thanks for the additional ideas.
Gekko, thanks for the clarification. I'm going to have to read that one a few more times to make sure I understand it all.

Also, Gekko I owe you an apology. This is your thread, started by you with a wonderful clip. It caught my attention because it's colorful and interesting and that led me to start asking questions that led away from your original post. So for that I am sorry.

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: Amoeba (video)

#16 Post by gekko » Fri Oct 24, 2014 8:10 am

Hi lilewis,
No apology needed. I enjoy and learn from discussions on this forum, and I believe the discussion in this thread that you have contributed to has added considerable value and interest to it, so thank you!

User avatar
vasselle
Posts: 2763
Joined: Sat Oct 25, 2014 5:32 pm
Location: France

Re: Amoeba (video)

#17 Post by vasselle » Mon Oct 27, 2014 4:11 am

Bonjour Gekko
Je n'avais pas vue cette vidéo
Elle est splendide
Merci pour le partage
Cordialement seb
Microscope Leitz Laborlux k
Boitier EOS 1200D + EOS 1100D

Post Reply