Macro to the extreme

[matroskin]

Nikon small world - http://www.nikonsmallworld.com/gallery/year/2010/11

Amazing stuff.

[MichaelChang]

<p>They sure are amazing; too bad we won't be able to make any of those images with the photography gear we have. Confocal microscopy is really expensive.<br>

[<a href="http://www.microscopyu.com/articles/confocal/confocalintrobasics.html">Link</a>]</p>

[graham_thompson1]

<p>Thanks for the link. Saw it and thought I want to do this.<br>

Then Michaels link spoilt it all :-(<br>

I wonder if it can be done on a more limited scale but less expensive.<br>

Standard microscope with adaptor?</p>

[MichaelChang]

<p>Yes you can, Graham, but of course with limited resolution compared to Kostya's original post.<br>

[<a href="http://www.google.ca/#hl=en&&sa=X&ei=i_K2TLK1F4GBlAfsvKS8DA&ved=0CBgQBSgA&q=photomicroscopy&spell=1&fp=a7248a2e7a166964">Link</a>]</p>

[bob_sunley]

<p>Have a look here as well, complete with how to do it pages.</p>

<p>http://www.krebsmicro.com/</p>

<p> </p>

[joseph_wisniewski]

<p>Hey Michael, not all the "Small World" images shot with confocal scopes this year. That's the 11th place image. Have a look at the winners.</p>

<ol>

<li>Jonas King shot a mosquito heart at a very conservative 100x, with a fluorescence scope. </li>

<li>Hideo Ostuna shot a zebrafish at 20x on a confocal scope.</li>

<li>Oliver Braubach shot a zebrafish's ovaries at 250x on a confocal scope</li>

<li>Riccardo Taiariol shot a wasp nest at 10x with "extended depth of field" stereomicroscopy</li>

<li>Viktor Sykora shot a seed at 10x with a darkfield scope.</li>

</ol>

<p>Bob Sunley pointed to a link by Charlie Krebs. He's a past "Small World" winner, and he uses what a serious microscopist would consider (politely) to be cobbled together, obsolete gear.</p>

<p>The fourth picture is a piece of cake. I've done (in my opinion, any way) better work. Technically, it's a snap. 10x with "extended depth of field", that's just focus stacking. There's many ways to get high quality 10x magnification, and most of those ways are "friendly" to focus stacking.</p>

<ul>

<li>Riccardo used a stereo microscope. Any decent stereo microscope should have tolerable 10x lenses. Stereo microscopes are not expensive. You rack (turn in small amounts) the scope's focus control, and shoot images at the different focus depths, then put them together in the really nice, totally free, and open source "CombineZ" software.</li>

<li>I could do it on my own scope, which isn't stereo (but does have nice lenses). My scope came from the Bay of E, and initially cost about $600 (more on that later). It came with 3 excellent Nikon CF objectives, and camera mount it came with was more than good enough for that 10x shot. It's only 10x, so rack focus with either big turns of the fine knob, or small turns of the coarse. And again with CombineZ.</li>

<li>I could do it without a microscope, at all, using the old Nikon PB-4 bellows (about $200 on the bay). The Nikon 10x CF makes a decent lens on the bellows (another $200 on the bay) or I could use the 16mm Zeiss Luminar. Rack focus using either the rail knob or the rear standard knob on the PB-4 (rear standard is more confusing, but produces a better looking stack).</li>

<li>Heck, I've done award winning 10x work with a Nikon 20mm f2.8 reversed on the front of a 200mm f4, and the focus stack done by shifting the camera and lens as a unit with a Novoflex Castel-L geared focus rail. That's two lenses that I already had, a coupling ring I made for $10 by gluing a pair of Cokin P-rings together face to face, and a $220 focus rail. I could just as easily used an $80 Manfrotto rail. </li>

</ul>

<p>The fifth picture is a seed at 10x, with darkfield illumination. Darkfield is a microscopist's way of saying "rim lighting". Viktor probably shot it on a microscope of some sort. My own scope shifts easily between darkfield and bright field. I can also do bright field with the bellows or the coupled 20mm and 200mm, using LEDs for the darkfield illumination. If you know how to rim light with big lights in a studio, you can do the same thing with little LED lights on a macro bench. Did you catch the thread where I posted snowflake pictures. Those were done with the darkfield using LEDs. I actually used colored lights, and colored darkfield is called "Rhineberg illumination". But the fancy name means "round up 6 LED booklights, $1 each from the dollar store, and gel them to color with little bits of filter material from a free Rosco sample book".</p>

<p>Looking at the first picture, my problem isn't equipment. Even though I use similar "junk" to Charlie, I can shoot that number one shot. My Nikon Optiphot 88 is a decade or two out of date, and I think my UFX camera adapter is a 1975 model. But my objectives are Nikon CF, and dang sharp. The whole scope cost me $600 on the Bay of E. I added fluorescence to both the epi and trans illumination systems with creative improvisation and under $100 in parts. The UV source is seven UV LEDs (and I'm ordering some shorter wavelength ones), and either replaces the source under the substage condenser, or slides into the tube for the epi system in front of the original light source. A trio of B+W 415 filters provide the UV blocking for the eyepieces for me and for the projection eyepiece for the camera.</p>

<p>The reason I can't match the first picture is that I have no idea how to microscopically dissect a mosquito, and dye the heart so magnificently for fluorescence. Nor do I have the vision to imagine a mosquito heart could have been such an amazing shot. King's skill, both technically and artistically, radically exceeds mine. That shot can be taken with less than $1000 in equipment, but the skills are worth possibly hundreds of thousands of dollars.</p>

[graham_thompson1]

<p>I am starting to find this interesting.<br>

Need to know more! What skills you all have. I just wish I had part. Interests me enough to - as we say in England - give it a punt.<br>

(Try it out)</p>

[bob_sunley]

<p>Well go for it, you can learn the skills, just like everyone else has for the last century or so. :) Start with reading Charlie Krebs pages, and there is a microscope group on Yahoo for help as well.</p>

<p>http://tech.groups.yahoo.com/group/Microscope</p>

<p> </p>