Cranefly genitalia

flynet · Post by **flynet** » Sat Nov 08, 2014 2:30 am

I started taking photos through the microscope early in 2014. It took me 3 months to decide on the equipment. After a few weeks experimenting, I began to take shots of fly genitalia, which was my intention. I have been writing papers on flies of the family Anthomyiidae since 1961, but until now have been making drawings with a camera lucida.
The camera I bought was a Canon EOS 0011D, which allows images to be seen on a monitor. The microscope a simple monocular. Stacking programme is Helicon Lite.
This photo is of the male genitalia of Molophilus appendiculatus, a small cranefly.

ChrisR · Post by **ChrisR** » Sat Nov 08, 2014 3:55 am

Welcome to the forum!

We look foward to seeing more of your work.
Any further details of your equipment (Canon 1100D ?) / microscope or techniques would be of interest to many members.

flynet · Post by **flynet** » Sat Nov 08, 2014 5:35 am

Photo of set-up, Canon 1100D (sorry I quoted from memory), with Brunel monocular SP27. They supplied all the camera attachments for this microscope, and a Meiji stereo, and an Old Watson monocular I have. The EOS software and cables came with the camera.

As I knew little about photography before (and only a bit now!) I read a lot of stuff on the web, bought PSE12 and manual, and just experimented for a few months.
The most important lesson I learnt was 1. for genitalia: make a good preparation for a slide (temporary one in glycol jelly on a slide 2. good lighting. The SP27 has an excellent LED light with battery or mains. For top light I use two flexible lights as used with stereos. Cost of equipment ( camera, monocular SP27, Photoshop 12, was about £550, most on the camera, the SP27 was under £200. My technique is just to experiment until I get the result I want!

flynet · Post by **flynet** » Sat Nov 08, 2014 5:44 am

For some reason I get either no pictures appearing with the message, or then I get two! Any expanation, what am I doing wrong?

ChrisR · Post by **ChrisR** » Sat Nov 08, 2014 6:25 am

Er, no, not really, holler if it happens again. I've taken the liberty of removing the duplicated photo.

Thanks for the details Brunel SP27 LINK.

If you have any questions, fire away.
I'm wondering if those cerci(?) should be blue

flynet · Post by **flynet** » Sat Nov 08, 2014 8:33 am

This is a photo of an anthomyiid head, taken with top light. Just a test to see if it uploads OK.

Thanks for deleting the duplicate, I could not find a way to do it! Also for the Brunel link.

ChrisR · Post by **ChrisR** » Sat Nov 08, 2014 8:59 am

Hitting the Edit buton should show the original text, where the links to the images should be clear..

I think you're losing some sharpness somewhere.
Here's link to something familar(?) taken by a pro but with a humble reversed enlarger lens:
http://www.photomacrography.net/forum/v ... 1076#51076

Do you use Live View to shoot from, on your camera? That should reduce vibrations. Your Camera does have "EFSC". You may find more references to the camera under its other designation, the Canon (Digital Rebel) T3.

flynet · Post by **flynet** » Sat Nov 08, 2014 9:27 am

The photos in your link are certainly very well focussed. But are these macro photos, i.e. not taken through the microscope but with lens and on a track? I am not equipped to do this. Probably the optics are not as good on the SP27, as some macro lens. Better stacking will help. I often leave unimportant characters out of focus in order to emphasize the important ones for taxonomic information. This makes for a more useful photo, but a less perfect one for a photographer.

I adjust the camera controls on the screen and press the shutter with the mouse. One problem I have noticed is that sometimes when focussing down with the fine focus when stacking is that the image jumps ahead and I have to focus back. The focus mechanism has not moved at all. It may be that I should wait awhile until the light has stabilised? It seems an optical problem.

The fly genitalia in your link is an American species of Eutrichota. There are about 40 species in the Nearctic, but I can only eliminate about half of them from the structure of the aedeagus.

ChrisR · Post by **ChrisR** » Sat Nov 08, 2014 9:45 am

T'other way round, your microscope should beat the lens used in the link - it's just a 50mm lens designed for a 35mm film enlarger. You will find examples of "noname" microscope objectives beating enlarger lenses quite easily, at 4x upwards.
It nearly always boils down to the "Effective Aperture" of the optics, before their price. Objectives generally win on that, over camera/macro/enlarger lenses.

"EFSC" (Electronic Front Shutter Curtain, where the sensor turns on without the shutter having to open mechanically) is very important - do a Search in the forum. Your shutter vibrations - even if you're using Mirror Lock, will give you problems. On your camera I think you can invoke it by shooting from Live View.

One way to check if vibrations are hurting you, is to rig up some well diffused (eg a tube of plain paper) flash. I'd still use Mirror Lockup.

(You can't use EFSC with flash, by the way, but using flash it doesn't matter as much. There's more on this, but it'll get confusing..)

flynet · Post by **flynet** » Sat Nov 08, 2014 1:34 pm

Thanks for this info Chris. I don't know if EFSC is operating in live view on my camera. There was no mention of it in my operating instructions.

How can I use a "well diffused flash" in a tube of paper when the light is coming from the LED light in the base of the microscope? It would have to come from below the stage, and there is no mirror to reflect it upwards.

I will do some research and see if I can find if the sensor is opening without the shutter.

Maybe see you sometime down here in Dorset?

Charles Krebs · Post by **Charles Krebs** » Sat Nov 08, 2014 3:25 pm

I don't know if EFSC is operating in live view on my camera. There was no mention of it in my operating instructions.

EFSC is always "on" with this camera when using live view. If you are viewing and operating the camera from the Canon remote software then you will be OK in this regard.

One problem I have noticed is that sometimes when focussing down with the fine focus when stacking is that the image jumps ahead and I have to focus back. The focus mechanism has not moved at all. It may be that I should wait awhile until the light has stabilised? It seems an optical problem.

Not quite sure I understand your problem, but is sounds like the fine focus may be slipping a little after you release it. It does sound mechanical rather than "optical".

On the issue of resolution... double check the aperture setting on your condenser. If it is set too small you will get a more contrasty image but you will lose resolution. You can usually get the contrast where you want in post processing, but there's no way to retrieve detail that was never recorded. You will generally get the best image with a slight reduction in the condenser diaphragm, but is is common that people go too far. This is an extremely common reason for lower resolution than might be otherwise be obtained.

The blue coloration in parts of your your image is due to the non-Apo color correction of the objective. This is usually pretty easy to correct if there are no elements in the image that are really blue.

In PSE go to Hue/Saturation. In the "drop-down" box select "Blue" instead of the default "Master". Reduce the saturation with the "Saturation" slider, and perhaps darken a little with the "Lightness" slider. (If needed, there are ways to fine-tune the range of blues that will be affected by this procedure using the eyedroppers and/or color bar at the bottom of the Hue/Saturation dialog box).

Here's a small section of you first shot before and after this quick simple adjustment.

(BTW... normally we don't edit or re-post images unless the poster encourages it. So if you would like me to remove this "blue" correction just say so).

ChrisR · Post by **ChrisR** » Sat Nov 08, 2014 4:42 pm

How can I use a "well diffused flash" in a tube of paper

The fly head is "top light". That'll do for a comparison.

Even with EFSC there could be wobbles.

flynet · Post by **flynet** » Sun Nov 09, 2014 2:14 am

Many thanks Charles for your reply. You are quite right, I generally have set the aperture to its smallest, probably because I thought the image looked better through the eye-piece or on the monitor. I will check this, and also if there is any slip in the fine focussing. I did tighten up the screws a bit as it was visibly slipping originally when I received it.

I don't think I have paid much attention to the Hue/Saturation control, though I have used some of the others extensively, especially the Dodge tool. PSE facilities take a long time to discover!

Kind of you to edit my Molopilus photo.

Both of you have given me plenty to think about, so thanks. This forum is living up to its intentions to help newcomers like me.

rjlittlefield · Post by **rjlittlefield** » Sun Nov 09, 2014 1:01 pm

flynet wrote:One problem I have noticed is that sometimes when focussing down with the fine focus when stacking is that the image jumps ahead and I have to focus back.

Sometimes an effect like this can be caused by slight stickiness in the stage movement, combined with a choice of moving the stage downward to step focus.

Try moving the stage upward to shoot the stack. That way focus movement will be driven by the gear train, which can easily develop large forces. When you step downward, the focus movement is controlled by the gear train but it's actually driven only by gravity and springs.

--Rik

flynet · Post by **flynet** » Mon Nov 10, 2014 2:04 am

Excellent suggestion Rik! I will try this in the next couple of days and come back to this thread.

By the way I have no indication in the literature which came with the gear (and on the web) about the shutter speed. I have been using 200 and 400, and tried up to 800.