Diatoms. Was: Boosting bright field resolution with dichroic filters

Posted by Steffen Dietzel on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Boosting-bright-field-resolution-with-dichroic-filters-tp7583983p7583994.html

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Well, I once tried diatomaceous earth from the health food store, only
to find out that it came well ground, so it wasn't much use for
microscopy. Haven't tried again ever since. If somebody should know a
brand available in Germany that does work, I would be very interested,
would be a great resource for teaching.

Anyway, I was thinking more in the direction of a source of cultured,
well defined (dead) diatoms, that will avoid the trouble of finding a
specialist to identify the species. Like "Pleurosigma, 10 g, 50 Euro" or
the like.

I am afraid that transgenic diatoms available on Amazon won't be a
reality any time soon in Germany... But whatever they use for
photosynthesis is probably fluorescent anyway, as long as they are alive.

Steffen

Am 13.07.2015 um 14:26 schrieb George McNamara:

> Diatoms in bulk ... maybe in your (or a colleague's) kitty litter.
>
> Diatomaceous earth
> https://en.wikipedia.org/wiki/Diatomaceous_earth
>
> Diatomaceous earth consists of fossilized remains ofdiatoms
> <https://en.wikipedia.org/wiki/Diatom>, a type of hard-shelledalgae
> <https://en.wikipedia.org/wiki/Algae>. It is used as afiltration
> <https://en.wikipedia.org/wiki/Filtration>aid, mild abrasive in products
> including metal polishes andtoothpaste
> <https://en.wikipedia.org/wiki/Toothpaste>, mechanicalinsecticide
> <https://en.wikipedia.org/wiki/Insecticide>,absorbent
> <https://en.wikipedia.org/wiki/Absorption_%28chemistry%29>for liquids,
> matting agent for coatings, reinforcing filler in plastics and rubber,
> anti-block in plastic films, porous support for chemical catalysts,cat
> litter <https://en.wikipedia.org/wiki/Cat_litter>, activator inblood
> clotting <https://en.wikipedia.org/wiki/Blood_clotting>studies, a
> stabilizing component ofdynamite
> <https://en.wikipedia.org/wiki/Dynamite>, and athermal insulator
> <https://en.wikipedia.org/wiki/Thermal_insulation>.
>
> (I first learned of it from using kilograms in the filtration system of
> our backyard pool).
>
> Or
> http://www.amazon.com/Diatomaceous-Earth-Food-Grade-10/dp/B00025H2PY
>
>
>     Diatomaceous Earth Food Grade 10 Lb
>
> <http://www.amazon.com/Diatomaceous-Earth-Food-Grade-10/dp/B00025H2PY/ref=sr_1_1?ie=UTF8&qid=1436789646&sr=8-1&keywords=Diatomaceous+earth>
>
> byDiatomaceousEarth
> Diatomaceous earth (de) is made of tiny, fossilized diatoms (aquatic
> organisms) that accumulated over millennia in fresh water lakes. When
> mined and left untreated, this amazing product can be used in hundreds
> of different ways. However, not all diatomaceous earth is created equal.
> While most other manufacturer's hand scoop de into zip lock bags,
> diatomaceous earth brand de is professionally packaged using stainless
> steel equipment. Because of this process, you can be confident there are
> no contaminants in your food grade diatomaceous earth.
>
>
> Maybe you could repackage this from "food grade" to "microscope grade".
>
> Same order could include
>
> http://www.amazon.com/100g-Fluorescein-Powder-Chemical-Reagent/dp/B00FJ64S3U
>
>
>     100g of Fluorescein Powder, Chemical Reagent
>
> <http://www.amazon.com/100g-Fluorescein-Powder-Chemical-Reagent/dp/B00FJ64S3U/ref=sr_1_1?ie=UTF8&qid=1436789750&sr=8-1&keywords=fluorescein>
>
> byAldon Corp
> $44.95
> <http://www.amazon.com/100g-Fluorescein-Powder-Chemical-Reagent/dp/B00FJ64S3U/ref=sr_1_1?ie=UTF8&qid=1436789750&sr=8-1&keywords=fluorescein>+
> $6.74 shipping
>
>
>
>
> Or make transgenic diatoms (http://www.ncbi.nlm.nih.gov/pubmed/25297896
> ... centric diatoms sounds good for FFTs, and their abstract mentions
> firefly luciferase and Azami-Green GP genes), fuse their silica binding
> protein (Si-tag, see, for example,
> http://www.ncbi.nlm.nih.gov/pubmed/21277372 ... Ecoli L2 ribosomal
> protein and its 203-273 peptide apparently are SiBP's and has been fused
> to EGFP, http://www.ncbi.nlm.nih.gov/pubmed/22926644) to GFP, RFP,
> photoswitchable FP, iRFP, etcFP,
> and sell that/those on amazon.com
>
> enjoy,
>
> George
>
>
> On 7/13/2015 4:26 AM, Steffen Dietzel wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Dear all,
>>
>> two questions, first about the formula:
>>
>> I thought 0.61*lambda/(NA)  (Rayleigh) would be the appropriate
>> formula for self-luminous objects (dark field, fluorescence) while for
>> bright field the fitting variant of the Abbe formula should be
>> applied: lambda/NA-obj for central illumination, lambda/2NA for NA-obj
>> = NA cond and, in general, lambda/(NA-obj + NA cond), provided that
>> NA-obj is equal or bigger than NA-cond.
>>
>> It seems though, that others are happy with 1.2*lambda/(NA-obj + NA
>> cond). Where does the 1.2 (or 2*0.6)  in a bright field situation come
>> from? Would that be the paper by Hopkins and Barham (1950), that Mike
>> was referring to? Because I have no hope of understanding this paper
>> by reading through during this life (Lots of formulas my biologist's
>> brain is not flexible enough to adjust for).
>>
>> Second:
>> Is there a good source for diatoms that can be stained with
>> fluorescent dyes and self-mounted, as George suggested? I have a few
>> diatom slides for teaching (Pleurosigma), but they came already mounted.
>>
>> Steffen
>>
>>>
>>>
>>> --
>>> ------------------------------------------------------------
>>> Steffen Dietzel, PD Dr. rer. nat
>>> Ludwig-Maximilians-Universität München
>>> Walter-Brendel-Zentrum für experimentelle Medizin (WBex)
>>> Head of light microscopy
>>>
>>> Marchioninistr. 27
>>> D-81377 München
>>> Germany
>>
>
>
> --
>
>
>
> George McNamara, Ph.D.
> Single Cells Analyst
> L.J.N. Cooper Lab
> University of Texas M.D. Anderson Cancer Center
> Houston, TX 77054
> Tattletaleshttp://works.bepress.com/gmcnamara/42
>


--
------------------------------------------------------------
Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Walter-Brendel-Zentrum für experimentelle Medizin (WBex)
Head of light microscopy

Marchioninistr. 27
D-81377 München
Germany