Illumination technique comparison using the same Closterium

[75RR]

Thought some of the new members of the forum who are also new to microscopy might like to see this comparison.
It is not always clear when one is starting out in microscopy how they differ - having them all together should help sort that out.
Although there are several illumination techniques - DIC tends to get a lot of the attention.
I myself have been guilty of overusing DIC (an occupational hazard) when one finally gets hold of a DIC system.

These have been published in Microbehunter forum before, in 2016. Thought I would resurrect them.

The first 3 images: Plan 16x/0.35, 720µm, Brightfield, Oblique and Darkfield, each consists of 2 stacks of 3 to 4 images stitched in Photoshop (Due to its extremely large size)
The 4th image: Neofluar 25x/0.60 Ph2, 720µm, Phase, 3 stacks of 4 images stitched in Photoshop (Reduced in size to fit in with the first 3)

Brightfield, Oblique, Darkfield and Phase

[Hobbyst46]

75RR - this is a beautiful display of beautiful objects.

Especially, in my opinion, BF, COL and DF. Phase is especially prominent for transparent objects, and yours here is clearly visible in BF. However, for the purpose of demonstration, I like it a lot.

please continue posting DIC, this microscopy mode is not so widespread outside funded research, I think.

[75RR]

Thanks Hobbyst46.
It is not COL (regret not making one though) but standard oblique, obtained by offsetting the brightfield port.

[billbillt]

75RR,

Thanks for sharing...


BillT

[Francisco]

Very interesting comparative

[JimT]

Well done and very instructional.

IMHO oblique is most pleasing due to the 3D effect but the DIF reveals more detail - some of which could be due to the higher magnification/ resolution.

All are very good images.

[charlie g]

Fantastic desmid images, 75RR, thank you for sharing this microscopy. Were the terminal pole vesicles gypsum granules jittering/ almost 'tinkeling' as you observed these living desmids?

All these image-captures are rich in a sense of plants doing their best to shape and maintain our dear world. Bravo to our planets primary producers! Charlie guevara

[75RR]

Many thanks to Hobbyst46, billbillt, Francisco, JimT and charlie g.

Were the terminal pole vesicles gypsum granules jittering/ almost 'tinkeling' as you observed these living desmids?

Yes they were ... I would describe it as a cosmic dance ;)


Just to clarify for those who have not ventured into stacking yet.

2 stacks of 3 to 4 images stitched in Photoshop
and
3 stacks of 4 images stitched in Photoshop

means that I photographed the Dismid in either 2 or 3 'parts' (each part consisting of either 3 or 4 individual images) and stacked each part;
I then stitched those stacks together in order to assemble an image that was able to capture the detail that only a larger magnification objective can.
A sort of photographic quilt if you like.

Here is the full size Phase image taken with the Neofluar 25x/0.60 Ph2 of this Giant South American Desmid which measured 750µm from tip to tip.

[GaryB]

Very nice.
I only recently acquired a phase objective and I have to admit I'm having way more fun with it than I thought I would. Anyone who has yet to try phase, I highly recommend it. Gives a whole new insight.

[coominya]

Brightfield, Oblique, Darkfield and Phase

Nice side by side 75RR. To my eye the Oblique image contains the most detail with the least interference. It seems to have almost the same fine detail as the phase capture but without the halo obscuring the clear edges. The only thing that seems to be lost is the nucleus and perhaps that small hole at the tip of the closterium.