[Pollinator] Extracting pollen from ethanol

[Asya Robertshaw]

Dear John Purdy,

Thank you so much for your email! Those are excellent questions. I've
managed to successfully get the pollen out of a few of my samples (using
the more time-consuming methods I described in my original email), and
mounted them on slides using the basic fuchsin jelly technique. I've
examined the slides under the microscope and centrifuging my samples didn't
seem to make pollen too clumped or covered with other materials. I'm able
to clearly see separate pollen grains and identify them using the pollen
library that I created last year.

Thank you so much for your suggestions on how to evaporate the ethanol
quicker. I'll definitely look into each of them!

Asya


On Mon, Aug 25, 2014 at 12:40 PM, John Purdy <johnrpurdy at gmail.com> wrote:

> Does the centrifuge cause pollen to cake and be more difficult to work
> with?
> Similarly does the process of drying down deposit any material that was
> dissolved in the ethanol onto the pollen grains making them harder to work
> with?
>
> There are 3 approaches to consider:
> 1) filtration through a 5 micron filter - there are filter cassettes that
> fit onto a syringe or you can use vacuum to draw the liquid through.
> 2) You can use a vacuum chamber that takes multiple samples simultaneously
> and dries down the solvent.
> 3) you can set the samples in a tray and use a gentle stream of air or
> nitrogen gas to speed up the evaporation. I think the apparatus with
> multiple air jets can be purchased.
>
>
>
> John Purdy PhD
>
>
>
>
> On Sat, Aug 23, 2014 at 3:53 PM, Asya Robertshaw <
> asya.robertshaw at gmail.com> wrote:
>
>> Hi all,
>>
>> I'm a graduate student in pollination biology and as part of my PhD
>> dissertation I am trying to identify pollen grains on insects' bodies. I've
>> collected a number of pollinators over the years that were stored in glass
>> vials in 70% ethanol until they were pinned. Now I am trying to extract the
>> pollen that was left behind in the ethanol when the insects were removed.
>> Does anyone have any suggestions for how to do that efficiently? I've tried
>> to simply let the ethanol evaporate from the vials, but that takes a very
>> long time. I've also tried centrifuging the solution, which works great.
>> However, because of the large volume of ethanol in each vial, I need to
>> transfer it to at least 7-8 microcentrifuge tubes, which is also not very
>> efficient considering how many total samples I have. If anyone has any
>> other methods or tips for getting the pollen out of the ethanol solution, I
>> would greatly appreciate it!
>>
>> Thanks!
>> Asya
>>
>> --
>> Asya Robertshaw
>> Doctoral Candidate
>> Purdue University
>> Dept of Botany & Plant Pathology
>> 915 W State Street
>> West Lafayette, IN 47907
>>
>> _______________________________________________
>> Pollinator mailing list
>> Pollinator at lists.sonic.net
>> https://lists.sonic.net/mailman/listinfo/pollinator
>>
>>
>


-- 
Asya Robertshaw
Doctoral Candidate
Purdue University
Dept of Botany & Plant Pathology
915 W State Street
West Lafayette, IN 47907
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