Doing Diatoms a Different Way

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billbillt
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Re: Doing Diatoms a Different Way

#61 Post by billbillt » Sat Dec 24, 2016 6:43 pm

Hi Charles,

Thanks for sharing more of your wonderful diatom photos,,

BillT

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rnabholz
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Re: Doing Diatoms a Different Way

#62 Post by rnabholz » Sat Dec 24, 2016 7:39 pm

Hey Charles,

Looks like you went right to work!

I am going to have to watch for the Surirella spiralis that would be a new one to me.

I am a bit alarmed about the web you describe. I will need to check my remaining sample to see if it is present. Please let me have any other observations that you make about it.

Keep the posts coming.

Rod

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Re: Doing Diatoms a Different Way

#63 Post by Charles » Sat Dec 24, 2016 9:03 pm

Thank you Bill and Rod,

Here is a picture of the Surirella spiralis. You can see better ones if you google it.
Surirella spiralis.jpg
Surirella spiralis.jpg (30.07 KiB) Viewed 5504 times
I can't explain the film. It is clinging to everything. I dry the strew slides open on the stirrer/heater and tried drying it hotter but the film is everywhere. Here is a picture of it in the strew where it is more concentrated. You can see the centric in the center encased in it as well as the Surirella on the left.
Strew Film2.jpg
Strew Film2.jpg (75.64 KiB) Viewed 5504 times

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KurtM
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Re: Doing Diatoms a Different Way

#64 Post by KurtM » Sun Dec 25, 2016 3:41 am

Sure that isn't Campylodiscus?
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Re: Doing Diatoms a Different Way

#65 Post by Charles » Sun Dec 25, 2016 1:11 pm

Kurt, I think you are right. It looks more like a Campylodiscus than a Surirella spiralis. Thank you!

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Re: Doing Diatoms a Different Way

#66 Post by Charles » Sun Dec 25, 2016 5:58 pm

So I cleaned the sample by boiling in HCL twice. The first 2 hr boil in HCL, I noticed a lot of the film floated to the top and was not dissolved by the HCL. I siphoned off the top third of HCL with most of the film and refilled with fresh HCL and boiled again for 2 hrs. More film floated to the top and this was also siphoned off and then did six water changes. And this is the strew now much cleaner:
Strew after HCL2.jpg
Strew after HCL2.jpg (41.63 KiB) Viewed 5482 times
Merry Christmas to you all!

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rnabholz
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Re: Doing Diatoms a Different Way

#67 Post by rnabholz » Mon Dec 26, 2016 4:30 pm

Wow, that must be some tough stuff. Sorry for the extra trouble.

I will be taking a look at the remainder of the sample that I have here. I am curious if it is something that is growing in the stored samples - hope not.

Looks good after your efforts.

Rod

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Re: Doing Diatoms a Different Way

#68 Post by rnabholz » Mon Dec 26, 2016 9:49 pm

Hey Charles,

I went back and took a look at the remaining vials of the Lake Michigan samples, and I can see the film you report with the naked eye.

So needless to say, I took a look at the other samples from other sources that I gathered this season. I cannot see any issues with them, at least obvious ones visible with the naked eye, so that is a bit of a relief. It would been crushing to have all of that effort go to waste.

If you come across anything like it in the other samples, let me know please.

Thanks

Rod

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Re: Doing Diatoms a Different Way

#69 Post by zzffnn » Tue Dec 27, 2016 12:31 am

Rod,

Your sample is NOT going to waste or being contaminated by microbes.

Most growing stuff (microbes) can be digested by muriatic acid. Yours is not.

Many small particles, such as diatom fragments or plant fiber pieces, have natural tendency to aggregate, forming a film. Diatom fragments and some plant fiber cannot be digested well by muriatic acid.

Edit:
I have heard people (Rene?) keeping cleaned diatoms in 10% muriatic acid. That increased ionic strength probably keeps small particles apart, as well as inhibit microbe growth.

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Re: Doing Diatoms a Different Way

#70 Post by Charles » Tue Dec 27, 2016 1:23 am

rnabholz wrote:Wow, that must be some tough stuff. Sorry for the extra trouble.

I will be taking a look at the remainder of the sample that I have here. I am curious if it is something that is growing in the stored samples - hope not.

Looks good after your efforts.

Rod
Rod, No worries. I was going to run the samples through HCL anyway. I don't think it was anything organic since HCL would have dissolved them. Whatever it is, it is as tough as the diatoms!

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Re: Doing Diatoms a Different Way

#71 Post by rnabholz » Tue Dec 27, 2016 10:17 pm

zzffnn wrote:Rod,

Your sample is NOT going to waste or being contaminated by microbes.

Most growing stuff (microbes) can be digested by muriatic acid. Yours is not.

Many small particles, such as diatom fragments or plant fiber pieces, have natural tendency to aggregate, forming a film. Diatom fragments and some plant fiber cannot be digested well by muriatic acid.

Edit:
I have heard people (Rene?) keeping cleaned diatoms in 10% muriatic acid. That increased ionic strength probably keeps small particles apart, as well as inhibit microbe growth.

Thanks zz, Glad to hear it is not likely something growing after the fact.

One way or another you guys are gonna drag me into acid.....that sounds more ominous that I intended it to be. ;^)

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Re: Doing Diatoms a Different Way

#72 Post by billben74 » Thu Dec 29, 2016 1:01 am

Just wanted to say what a cool thread this is. I'm sure the whole forum will be reading this one.
Great work Charles. You could be inspiring a wave of future diatomists.

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Re: Doing Diatoms a Different Way

#73 Post by Charles » Thu Dec 29, 2016 1:10 pm

Thank you billben!

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Re: Doing Diatoms a Different Way

#74 Post by billbillt » Thu Dec 29, 2016 4:40 pm

billben74 wrote:Just wanted to say what a cool thread this is. I'm sure the whole forum will be reading this one.
Great work Charles. You could be inspiring a wave of future diatomists.
I agree!.. It will spark more interest in this interesting subject....

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Re: Doing Diatoms a Different Way

#75 Post by Charles » Wed Jan 11, 2017 5:05 pm

I've been down sick for the last couple of weeks but have been back at it the last few days.

One thing I noticed is my discard wash water in a beaker (which had been setting for a couple of weeks) had settled some film on the bottom of the beaker. So, wondering what it was, I siphoned off most of the water using a 50ml syringe and large bore needle (this was taken from a turkey injection system), I put the bottom scum into a tube added distilled water and put it on a couple of slides. This is the wash water I siphoned off samples which I was processing from the three samples James River, Chesapeake Bay and Virginia Beach. What was on the slide from the wash water was amazing. The reject water had almost every form which I had seen in the three samples, and there were some very large forms. I did the same for the sandy bottom sediment of centrifuge tubes trying to separate diatoms from the boulders. Doesn't make pretty strew slides but I was able to picked them off the slide onto a storage slide, which I will post photos of soon.

So, don't throw out that was wash water until you see what is in there.

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Re: Doing Diatoms a Different Way

#76 Post by rnabholz » Wed Jan 11, 2017 7:22 pm

You know that you are going to make me lie awake at night don't you?

I always hesitate to discard any of that kind of material, completely sure that it must contain the coolest, rarest, previously undiscovered diatoms. Now I can't ever get rid of any of it..... Thanks a lot Charles. ;^)

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Re: Doing Diatoms a Different Way

#77 Post by Charles » Thu Jan 12, 2017 12:55 pm

rnabholz wrote:You know that you are going to make me lie awake at night don't you?

I always hesitate to discard any of that kind of material, completely sure that it must contain the coolest, rarest, previously undiscovered diatoms. Now I can't ever get rid of any of it..... Thanks a lot Charles. ;^)
Then I'm going to realize your worst fears. As well as finding almost every species from the regular strew, I found at least three species which I didn't find in the regular strews. Like a couple of large different variety of Actinoptychus, three different varieties of Triceratium (encluding very large - over 100nm forms), as well as a very large Auliscus and other large centrics.

I'll try to throw some pictures up later today and you will be amazed!

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Re: Doing Diatoms a Different Way

#78 Post by rnabholz » Thu Jan 12, 2017 1:56 pm

I am really surprised that you found such large forms in the mix. Logic would argue that the large forms would have precipitated out before you drew the water off.

I wonder why it seems that they were floating about?

Read you post through blood shot eyes having been awake all night.... :^)

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Re: Doing Diatoms a Different Way

#79 Post by Charles » Fri Jan 13, 2017 12:59 am

Some of the large forms came from the 'sand wash water', which was being thrown out and some from just 'film wash water'. But even the 'film water' had large forms which really surprised me. Maybe the siphon was getting too near the bottom where the diatoms had settled and sucked some up.

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Re: Doing Diatoms a Different Way

#80 Post by KurtM » Fri Jan 13, 2017 1:03 am

Pictures! Pictures! :ugeek:
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Re: Doing Diatoms a Different Way

#81 Post by Charles » Fri Jan 13, 2017 1:10 am

Here are some pictures of the 'Sand waste water' which was being thrown out. At first I thought I would line them up (centrics to the left and pennates to the right) to see how many different forms I could find in the waste water, but then I liked how it looked and will probably do all my storage slides the same way. These were all taken on the B&L StereoZoom7 at about 40X (2.5x zoom and 15x eyepiece).

First the forms from the James River sample sand sample:
Storeage Rocks JR.JPG
Storeage Rocks JR.JPG (58.08 KiB) Viewed 5315 times
This is from the Buckroe Beach sand sample:
LStoreage Rocks BB.jpg
LStoreage Rocks BB.jpg (60.54 KiB) Viewed 5316 times
And the third is from Virginia Beach sand sample:
LStoreage Rocks VB.jpg
LStoreage Rocks VB.jpg (57.95 KiB) Viewed 5320 times
Last edited by Charles on Fri Jan 13, 2017 1:39 am, edited 3 times in total.

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Re: Doing Diatoms a Different Way

#82 Post by Charles » Fri Jan 13, 2017 1:28 am

Here are the forms taken from the bottom film from the beaker where I siphoned the rinse water into and it set for about 3 days. When I siphon, I try to get very near the bottom so some of the diatoms may have been sucked up and out during the siphon although I try to be real careful. The rinse water from all three sources were collected in the same beaker so the forms are from all three areas...James River, Buckroe Beach and Virginia Beach. Every form from my regular strew is in there except I didn't find any Hydrosera but I did find forms I hadn't found in the regular strew slides.

Here are pictures of the waste water film diatoms. I tried to make the arrangement it a bit more aesthetic and this probably how I will do my storage slides from now on. It gives a good visual the different kinds and size of forms.

Waste water film diatoms full view at about 2.5X setting on the B&L StereoZoom7:
LStoreage Film.jpg
LStoreage Film.jpg (51.57 KiB) Viewed 5317 times
Left half view close up at about 5X setting:
LStoreage Film2.jpg
LStoreage Film2.jpg (51.13 KiB) Viewed 5317 times
Right half view close up at about 5X setting:
LStoreage Film3.jpg
LStoreage Film3.jpg (47.7 KiB) Viewed 5317 times
Second ring on this storage slide where I initially put all the forms, which I then clean and pick from. These have not been picked through yet, but I will put most of these onto the final arrangement line eventually...and there are six more strews to go through:
LStoreage Film Extra.jpg
LStoreage Film Extra.jpg (65.14 KiB) Viewed 5317 times
Gave me a wake up call to be more careful in cleaning! :)

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Re: Doing Diatoms a Different Way

#83 Post by Charles » Fri Jan 13, 2017 1:42 am

KurtM wrote:Pictures! Pictures! :ugeek:
Pictures are up! :)

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Re: Doing Diatoms a Different Way

#84 Post by zzffnn » Fri Jan 13, 2017 1:50 am

Very cool! Thank you for sharing!
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Re: Doing Diatoms a Different Way

#85 Post by KurtM » Fri Jan 13, 2017 4:34 am

Well, that just fairly blows me away - absolutely fantastic work, Charles! It's inspired, and inspiring! 8-) 8-) 8-)
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Re: Doing Diatoms a Different Way

#86 Post by btschumy » Fri Jan 13, 2017 5:46 am

Charles,

Great pictures!

How do you go about lining the diatom's up like that? Could you describe your technique? I assume you use some sort of micro pipette and don't pick them up with tiny forceps :lol:
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Re: Doing Diatoms a Different Way

#87 Post by exmarine » Fri Jan 13, 2017 11:53 am

Yes absolutely fantastic work, I hope you don't mind but I have downloaded some of your photos for my reference. Thank you they are great.
Thank you :shock:
Best regards
exmarine :x

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uses Watson Stereo 1960 ish.

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Re: Doing Diatoms a Different Way

#88 Post by rnabholz » Fri Jan 13, 2017 2:03 pm

Charles!

All I can say is wow. Amazing work, so precisely done, with so much dedication. Just wonderful.

Now after viewing the work of a fine Renaissance master painter, I return to my work that looks a little bit like monkey got hold of a paintbrush.... ;^)

Great post

Rod

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Re: Doing Diatoms a Different Way

#89 Post by Charles » Sun Jan 15, 2017 5:11 am

Thanks to you all!

btschumy, It is just some micro-manipulation with a drawn needle. If you go to the first page of this post, you will see my set up, tools and technique.

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Re: Doing Diatoms a Different Way

#90 Post by Charles » Wed Jan 25, 2017 2:31 pm

Just a quick follow-up. I conducted some a test on how fast the diatoms settle.

I put about 1 ml of diatoms into a 15ml test tube and mixed it well with distilled water and let it settle. At different intervals I took a sample from the top and put three drops onto a slide with the following results:

1 minute: Lots of diatoms present
2 minute: Diatoms still present
3 minute: Just a few diatoms present
4 minute: Very few diatoms present
5 minute: Almost no diatoms present

During the cleaning process and water changes, I usually let it settle about 2 hrs. So the diatoms I'm seeing in the saved discard water from the water changes most like came from me suctioning too close to the bottom near where the diatoms have settled. And these are being sucked up by the force of the suction.

There's no getting around the sand sediment because some diatoms (especially the larger/denser forms) sink almost as fast as the sand particles. Not a problem for making strews for picking, but would be a problem for those making strews for permanent mounts.

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