Good afternoon,

Dear Greg,

your question starts with "recipe for 1% Acetocarmine?" followed by "what is the best way to store the solution, and how long is it viable?"

Knowing that it might be of course very easy to get a practical and tested recipe with additional comments from a colleague out in the wild (which is/has been left open for now 2 days, 6 views) I wonder about your not having "googled" before (or did you?), because:

Google search for < Acetocarmine AND pollen AND STAINING > yields approx. 15,000 results, for

< Acetocarmin staining solution > = 29.400 results and Google Search for:

< acetocarmine stain > yields approx. 27,500 results, at least the first 20-30 results of each search containing valid information with relation to your question.

You don't say whether you already have a suited acetocarmine solution (which can be purchased too

[e.g. http://www.chemicalbook.com/ChemicalProductProperty_EN_CB91876528.htm ; ProChem, Inc., [email protected], USA # CB91876528; or http://www.espchemicals.com/Aceto-Carmine-Stain-Schneider-Aqueous-Solution-C1964.aspx: # A1343-02 :

Aceto-Carmine Stain,(Schneider),Aqueous Solution (1 x 100 ml); or http://www.scholarchemistry.com/groups.php?id=15003 : # 9700004 - Aceto-Carmine Solution, 2% (aqueous), 100 mL Bottle, etc.] ) so it might not be that difficult to get or produce a 1% solution, see the following URL) or need also company order details for the Carmine dye.

The best recipe to be found very rapidly was:

http://www.k-state.edu/wgrc/Protocols/Cytogenetics/acetocarmine.html

Concerning storage and durability / viability of the solution (related +/- to CI 75470, CI Natural red 4) out of CONN's Biological Stains, 10th Ed, p.372ff (Carminic acid, Carmine and Cochineal; HOROBIN & KIERNAN Eds, BIOS Scientific Publishers Ltd, Oxford UK) I found:

"Stability of stock and staining solutions vary widely. Alkaline solutions are unstable. Light fastness is good."

Reference in that standard book: Acetocarmine in Botanical material....see

RUZIN SE,(1999) Plant Microtechnique and Microscopy. Oxford University Press, New York.

Personal note: since the solution contains 45% acid it is recommended to store solution in tightly closed dark screw cap bottle, in cool, dry, well-ventilated place away from incompatible materials, usually in a cabinet for corrosive chemicals. Use gloves and personal protection if appropriate. Wash thoroughly after handling.

There are several versions of this stain.

An iron containing stain is often used because of the darker bluish-red color produced cf. http://www.thelabrat.com/protocols/Aceto-Carmine.shtml

or at http://www.changbioscience.com/protocols/recipe/Acetocarminestainingsolution1.htm you'll find:

Acetocarmine staining solution (1%)

Carmine 1 gram (g)

Glacial acetic acid (45%) 100 millilitre (ml)

Ferric Chloride (FeCl2*6H2O) (10%) Optional

5 millilitre (ml)

Total volume 100 millilitre (ml)

Note: Add carmine powder to boiling 45% glacial acetic acid, cool rapidly, and then filter into a dark glass. Staining can be intensified by adding ferric chloride (FeCl2.6H2O). Caution: the stain stains clothes.

Hoping this would be of help to get started...

best regards,

Wolfgang MUSS, Salzburg-Austria

Good afternoon,

Dear Greg,

your question starts with "recipe for 1% Acetocarmine?" followed by "what is the best way to store the solution, and how long is it viable?"

Knowing that it might be of course very easy to get a practical and tested recipe with additional comments from a colleague out in the wild (which is/has been left open for now 2 days, 6 views) I wonder about your not having "googled" before (or did you?), because:

Google search for < Acetocarmine AND pollen AND STAINING > yields approx. 15,000 results, for

< Acetocarmin staining solution > = 29.400 results and Google Search for:

< acetocarmine stain > yields approx. 27,500 results, at least the first 20-30 results of each search containing valid information with relation to your question.

You don't say whether you already have a suited acetocarmine solution (which can be purchased too

[e.g. http://www.chemicalbook.com/ChemicalProductProperty_EN_CB91876528.htm ; ProChem, Inc., [email protected], USA # CB91876528; or http://www.espchemicals.com/Aceto-Carmine-Stain-Schneider-Aqueous-Solution-C1964.aspx: # A1343-02 :

Aceto-Carmine Stain,(Schneider),Aqueous Solution (1 x 100 ml); or http://www.scholarchemistry.com/groups.php?id=15003 : # 9700004 - Aceto-Carmine Solution, 2% (aqueous), 100 mL Bottle, etc.] ) so it might not be that difficult to get or produce a 1% solution, see the following URL) or need also company order details for the Carmine dye.

The best recipe to be found very rapidly was:

http://www.k-state.edu/wgrc/Protocols/Cytogenetics/acetocarmine.html

Concerning storage and durability / viability of the solution (related +/- to CI 75470, CI Natural red 4) out of CONN's Biological Stains, 10th Ed, p.372ff (Carminic acid, Carmine and Cochineal; HOROBIN & KIERNAN Eds, BIOS Scientific Publishers Ltd, Oxford UK) I found:

"Stability of stock and staining solutions vary widely. Alkaline solutions are unstable. Light fastness is good."

Reference in that standard book: Acetocarmine in Botanical material....see

RUZIN SE,(1999) Plant Microtechnique and Microscopy. Oxford University Press, New York.

Personal note: since the solution contains 45% acid it is recommended to store solution in tightly closed dark screw cap bottle, in cool, dry, well-ventilated place away from incompatible materials, usually in a cabinet for corrosive chemicals. Use gloves and personal protection if appropriate. Wash thoroughly after handling.

There are several versions of this stain.

An iron containing stain is often used because of the darker bluish-red color produced cf. http://www.thelabrat.com/protocols/Aceto-Carmine.shtml

or at http://www.changbioscience.com/protocols/recipe/Acetocarminestainingsolution1.htm you'll find:

Acetocarmine staining solution (1%)

Carmine 1 gram (g)

Glacial acetic acid (45%) 100 millilitre (ml)

Ferric Chloride (FeCl2*6H2O) (10%) Optional

5 millilitre (ml)

Total volume 100 millilitre (ml)

Note: Add carmine powder to boiling 45% glacial acetic acid, cool rapidly, and then filter into a dark glass. Staining can be intensified by adding ferric chloride (FeCl2.6H2O). Caution: the stain stains clothes.

Hoping this would be of help to get started...

best regards,

Wolfgang MUSS, Salzburg-Austria

I did google it, but I was fishing to see if someone had a tested recipe that they recommended. Prior to this post, I had conferred with other colleagues which recommended different recipes. I was hoping to come up with some kind of consensus as I am new to using this stain. The link at the bottom were helpful.

Thanks.

Dear Greg, you're welcome,

I'm not quite confident about presence of plant researchers at Researchgate (especially dealing with stains for plants and their components...wish you the best for a successful staining!. As I have seen too, there are some articles concerning specificity of the stain (e..g Alexander's and others).

Good luck, Wolfgang

Probably my help is outdated, but for other fellows who stumble in this post, I found these sources or papers useful specifically for pollen viability using Acetocarmine.

http://www.shigen.nig.ac.jp/wheat/wis/No26/p1/1.html

http://tru.uni-sz.bg/tsj/Vol7No1_2009/Sohail1.pdf

http://www.le.ac.uk/bl/phh4/openpubs/openpubs/SPR_HH_pollen_stigma_crestiLR.pdf

http://www.hindawi.com/journals/isrn/2013/207434/

Greetings from M A L T A

As per my littel experience in sunflower crop. I ma herewith sending two ways to make 1% Acetocarmine solution for pollen study.

How to prepare Acetocarmine staining solution (1%)

Carmine: 1gram (g)

Glacial acetic acid (45%):100 millilitre (ml)

Ferric Chloride (FeCl2*6H2O) (10%) Optional: 5 millilitre (ml)

Total volume: 100 millilitre (ml)

Note: Add carmine powder to boiling 45% glacial acetic acid, cool rapidly, and then filter into a dark glass. Staining can be intensified by adding ferric chloride (FeCl2.6H2O).

There are two types of version presented below.

Non-iron version

Iron containing version

If you are facing any problem to making 1% Acetocarmine solution directly you can purchase from market.

Second question regarding how to store : Simly you can keep in normal room otherwise you can keep in freeze also. Same stain you can use atleast a year without any problem you will get perfect result. But some time results vary from crop to crop. I am using a year old acetocarmine solution for pollen study and getting excellent result.

@ Hari Prakash Meena: some minute ago I found your most recent reply which has been posted 7 months ago...(Unfortunately I found the e-mail notification right now).

I recommended your reply [recommendation 3] too - because you gave solutions / recipes - but then found that you gave data which are contained in bibliographic sources you did NOT cite (see my Reply # 00,above):

Quote: "...... or at http://www.changbioscience.com/protocols/recipe/Acetocarminestainingsolution1.htm you'll find:

Acetocarmine staining solution (1%)

Carmine 1 gram (g)

Glacial acetic acid (45%) 100 millilitre (ml)

Ferric Chloride (FeCl2*6H2O) (10%) Optional

5 millilitre (ml)

Total volume 100 millilitre (ml)

Note: Add carmine powder to boiling 45% glacial acetic acid, cool rapidly, and then filter into a dark glass. Staining can be intensified by adding ferric chloride (FeCl2.6H2O). Caution: the stain stains clothes.

Hoping this would be of help to get started...

best regards,

Wolfgang MUSS, Salzburg-Austria" End of quote

which is definitely the same you posted long after the first answers have been given.

The second part of your post is taken +/- word by word from several sources:

e.g.,

http://groveseducation.weebly.com/uploads/4/7/4/5/4745137/onion_root_tip_preparation.doc

The only sentences you left out at the end (and copied &pasted to Point 1) are those regarding working safety with acids....

The whole text is found also in:

www.dave-cushman.net/bee/microform.html,

http://www.thelabrat.com/protocols/Aceto-Carmine.shtml

and perhaps other documents in the web, too.

It is 'policy' (not politeness!) of this forum / ResearchGate Platform that academics or those who are going to become academics, researchers, teachers, professors and investigators are expected to observe the rules of GSP(= „Good Scientific Practice“) which means that external sources (sources not created by oneself) have to be /are to be mentioned,

otherwise - even in this forum - you / they are committing 'plagiarism'.

Therefore I retract my previously given recommendation to your reply # 005.

With no compliments, W.M.

Thank you sir. I think it may be because of some issues related to website or it may be due to some other reasons.

Dear Dr Stephen Mifsud thank you for sharing these resources.