[Nhcoll-l] comparative quality cryogenic vials

Thu Aug 20 10:55:08 EDT 2020

Greg

We took all of those things into account when selecting our long-term storage tubes.  The tubes that we selected are cryo-safe (vapor NOT liquid), externally threaded (prevents thread coming into contact with large chunks of tissue and also prevents possible contamination), and are the right size to fit into an 81 tube box (they fit well and do not rattle in the box which some of the thinner tubes do.  We are also happy with losing extra capacity for safety of tissues).

In terms of evaporation, we have had no issues with evaporation in the field or returning from the field but we also routinely decant off all liquid before placing samples in the LN2 dewars for two reasons – to prevent damage to DNA from ethanol over time (it will be arrested somewhat due to low temp but still happens) and to ensure access to the tissue sample in the tube (95% ethanol and RNALater both freeze at LN2 temperatures and so you have to thaw the liquid before being able to access the tissue which may degrade DNA quality over time with multiple freeze:thaw cycles).

There is some info on tissue collections on the SPNHC wiki:  https://spnhc.biowikifarm.net/wiki/Tissue_Sample_Collection but I suspect that the vial selection section needs updating

Andy

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Andy Bentley
Ichthyology Collection Manager
University of Kansas
Biodiversity Institute
Dyche Hall
1345 Jayhawk Boulevard
Lawrence, KS, 66045-7561
USA

Tel: (785) 864-3863
Fax: (785) 864-5335
Email: abentley at ku.edu<mailto:abentley at ku.edu>
http://ichthyology.biodiversity.ku.edu<http://ichthyology.biodiversity.ku.edu/>

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From: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu> on behalf of Greg Watkins-Colwell <gregory.watkins-colwell at yale.edu>
Date: Thursday, August 20, 2020 at 8:22 AM
To: "nhcoll-l at mailman.yale.edu" <nhcoll-l at mailman.yale.edu>
Subject: Re: [Nhcoll-l] comparative quality cryogenic vials

Has anybody ever written a paper on this?  It would sure be helpful.  Each of the tubes that Andy and Dirk listed are the opposite of what I was told to use when we migrated to LN2 (internal thread with the silicon O-ring..).

There’s also an issue I’ve found with some tubes not fitting properly in the boxes we use in the LN2 tanks.  Some tubes are too narrow and rattle around.  Others are too tall and the lid won’t go on properly (nor will the box properly fit in the rack as a result).  So there are clearly a lot of factors.  When we migrated to LN2 we had a lot of things in external thread, black o-ring tubes and when asked if those tubes are okay to use in LN2, the vendor literally shrugged and said… very slowly… “prob-ab-ly?”… so we transferred everything.  Very time-consuming!

A nice paper detailing the pros and cons of each with a solid recommendation would be great.

On the issue of evaporation, we routinely decant off the liquid anyway before putting the tubes into LN2.  Also, many of our samples are frozen raw (especially in ORN and MAM).  How does tube morphology impact tissue samples in those cases?  Anybody have any data on that?

Greg

****************
Gregory J. Watkins-Colwell
Sr. Collection Manager, Herpetology and Ichthyology
Division of Vertebrate Zoology
Yale Peabody Museum of Natural History
Main Office: 203-432-3791;  West Campus: 203-737-7568; Fax 203-432-9277

Package shipping address:
Greg Watkins-Colwell
Division of Vertebrate Zoology
Yale Peabody Museum of Natural History
170–210 Whitney Avenue
New Haven, CT 06511 USA
203-432-3791
******************

From: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu> On Behalf Of Bentley, Andrew Charles
Sent: Wednesday, August 19, 2020 5:20 PM
To: nhcoll-l at mailman.yale.edu
Subject: Re: [Nhcoll-l] comparative quality cryogenic vials

John

We have been using these Nalgene tubes in our collection and for collecting in all environments for over 15 years and have had no issues with storage in 95% ethanol, RNALater or liquid nitrogen environments:  https://www.fishersci.com/shop/products/nalgene-general-long-term-storage-cryogenic-tubes/033377d<https://nam10.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.fishersci.com%2Fshop%2Fproducts%2Fnalgene-general-long-term-storage-cryogenic-tubes%2F033377d&data=02%7C01%7Cabentley%40ku.edu%7Cbcf5103bcb2d46bdbabd08d8450c153f%7C3c176536afe643f5b96636feabbe3c1a%7C0%7C1%7C637335265528151199&sdata=6IApZTN4%2F5bc8AqctU2HkCCBe0RehbCzubpjNrkRBo8%3D&reserved=0>

I agree with Dirk that whatever make you go with, externally threaded tubes with no O-ring gasket are the way to go.

Andy

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Andy Bentley
Ichthyology Collection Manager
University of Kansas
Biodiversity Institute
Dyche Hall
1345 Jayhawk Boulevard
Lawrence, KS, 66045-7561
USA

Tel: (785) 864-3863
Fax: (785) 864-5335
Email: abentley at ku.edu<mailto:abentley at ku.edu>
http://ichthyology.biodiversity.ku.edu<https://nam10.safelinks.protection.outlook.com/?url=http%3A%2F%2Fichthyology.biodiversity.ku.edu%2F&data=02%7C01%7Cabentley%40ku.edu%7Cbcf5103bcb2d46bdbabd08d8450c153f%7C3c176536afe643f5b96636feabbe3c1a%7C0%7C0%7C637335265528161195&sdata=D8JsyzBQvwBaUpoOqKvTbaAaY8OrtPZ3EU2bsnEmOds%3D&reserved=0>

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From: Nhcoll-l <nhcoll-l-bounces at mailman.yale.edu<mailto:nhcoll-l-bounces at mailman.yale.edu>> on behalf of Dirk Neumann <neumann at snsb.de<mailto:neumann at snsb.de>>
Reply-To: "neumann at snsb.de<mailto:neumann at snsb.de>" <neumann at snsb.de<mailto:neumann at snsb.de>>
Date: Wednesday, August 19, 2020 at 4:05 PM
To: "nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>" <nhcoll-l at mailman.yale.edu<mailto:nhcoll-l at mailman.yale.edu>>
Subject: Re: [Nhcoll-l] comparative quality cryogenic vials

Dear John,

I would prefer NUNC Store-it tubes (article 264300; https://www.thermofisher.com/order/catalog/product/264262#/264262<https://nam10.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.thermofisher.com%2Forder%2Fcatalog%2Fproduct%2F264262%23%2F264262&data=02%7C01%7Cabentley%40ku.edu%7Cbcf5103bcb2d46bdbabd08d8450c153f%7C3c176536afe643f5b96636feabbe3c1a%7C0%7C1%7C637335265528161195&sdata=hy7k71Bb7L9oAvewE%2FOdfZd8zb%2BP%2FxM8%2BnOvDpiHHPk%3D&reserved=0>)

We tested them against Sarsted and other brands in a warm water bath > 50°C and they were among the tightest ones; also, they DON'T have an silicone O-ring (which will fail after some time); we had to replace poor quality tubes twice so far and besides necessary replacement costs and time needed to transfer tissues the huge evaporation loss (> 50% in some tubes) surely affected the integrity of the tissues / DNA.

The NUNC-tubes have a flange and really need to be screwed down tight (which is a bit annoying), but we have them on storage since 2004, use them in the tropic in the field and their performance is excellent. Bit more expensive, but this slightly higher investment pays off in the long run.

Hope this helps,
with best wishes
Dirk

Am 17.08.2020 um 23:42 schrieb John Edwin Lattke Bravo:
Hello,

Does anybody have any thoughts or experience to share regarding the comparative merits of 2ml cryogenic vials by Corning, Sarstedt and Granier for long term storage of insects in ethanol? Is the quality amongst the three similar? Positive experiences? Negative one? Just needing some user experience to help decide what might deliever the best bang for the buck (or real, in my case).

Obrigado,

John Lattke

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