Illumination technique comparison using the same Closterium

Here you can discuss different microscopic techniques and illumination methods, such as Brightfield, Darkfield, Phase Contrast, DIC, Oblique illumination, etc.
Post Reply
Message
Author
User avatar
75RR
Posts: 6243
Joined: Sun Oct 12, 2014 2:34 am
Location: Estepona

Illumination technique comparison using the same Closterium

#1 Post by 75RR » Tue Mar 20, 2018 9:01 am

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
Image Image Image Image
Last edited by 75RR on Tue Mar 20, 2018 3:46 pm, edited 2 times in total.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

Hobbyst46
Posts: 2157
Joined: Mon Aug 21, 2017 9:02 pm

Re: Illumination technique comparison using the same Closterium

#2 Post by Hobbyst46 » Tue Mar 20, 2018 10:34 am

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.
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

User avatar
75RR
Posts: 6243
Joined: Sun Oct 12, 2014 2:34 am
Location: Estepona

Re: Illumination technique comparison using the same Closterium

#3 Post by 75RR » Tue Mar 20, 2018 2:39 pm

Thanks Hobbyst46.
It is not COL (regret not making one though) but standard oblique, obtained by offsetting the brightfield port.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

billbillt
Posts: 2885
Joined: Mon Nov 10, 2014 10:01 pm

Re: Illumination technique comparison using the same Closterium

#4 Post by billbillt » Tue Mar 20, 2018 3:32 pm

75RR,

Thanks for sharing...


BillT

Francisco
Posts: 327
Joined: Sun May 28, 2017 11:23 pm

Re: Illumination technique comparison using the same Closterium

#5 Post by Francisco » Tue Mar 20, 2018 5:35 pm

Very interesting comparative

JimT
Posts: 3247
Joined: Fri Oct 24, 2014 1:57 pm

Re: Illumination technique comparison using the same Closterium

#6 Post by JimT » Tue Mar 20, 2018 9:12 pm

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
Posts: 1401
Joined: Mon Oct 20, 2014 7:54 pm

Re: Illumination technique comparison using the same Closterium

#7 Post by charlie g » Wed Mar 21, 2018 2:01 am

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

User avatar
75RR
Posts: 6243
Joined: Sun Oct 12, 2014 2:34 am
Location: Estepona

Re: Illumination technique comparison using the same Closterium

#8 Post by 75RR » Wed Mar 21, 2018 6:14 am

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.

Image
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

GaryB
Posts: 77
Joined: Tue Sep 05, 2017 10:33 am

Re: Illumination technique comparison using the same Closterium

#9 Post by GaryB » Thu Mar 22, 2018 7:21 am

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.

User avatar
coominya
Posts: 279
Joined: Sun Mar 11, 2018 11:33 am
Location: Brisbane Aust

Re: Illumination technique comparison using the same Closterium

#10 Post by coominya » Fri Mar 30, 2018 9:18 am

75RR wrote:
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.

Post Reply