I was thinking of oblique-relief-phase contrast attempts so dug a strew slide of benthic marine diatoms, and searched for a large one among the many 20-30um frustules. These diatoms
I had collected about 1.5 year ago, gently cleaned with SDS+EDTA +3% H2O2 and mounted in NOA61 (not the best mountant but that is what I used then).
And for the first time, I noticed a Pleurosigma (tentative ID!). A whole undamaged 110um-long frustule. Lying among the smaller diatoms and organic remnants (those would have been destroyed had I used corrosive chemicals).MicroBob wrote:It is always astonishing, how long one can look at a strew slide and still found new diatom forms sponge needles and other interesting oblects.
I was very happy with it, so threw away the original purpose of the inspection and just acquired images under diverse illuminations, using mostly the 100X1.3 phase contrast (condenser not oiled). And the simple 100X1.25 achromat for semi-dark field. This objective and the darkfield Ultracondenser were both oiled.
The brightfield image creates a "shadow" along the top rim of the diatom. This artifact occurs in the specific plane of focus that I chose to sharpen as many dots as possible.
Some of the images obtained with a green interference filter, peak 543nm, bad width (at half height) 25nm. Those images required ~4-fold long exposures.
Images are mostly untouched except for resizing and stacking.
Comments are welcome.