[Histonet] Message for Mark Ray

Tue Oct 5 11:05:06 CDT 2004

Dear Mark,

Our mail server says your email address
darkdaym <@t> mindspring.com
does not exist, so I'm sending the message to Histonet
instead, with your name in the subject line.

Yes, safranine has an e and so does rhodamine.
Acid fuchsine gets an e because although it's
an anionic dye it's derived from an organic base
by addition of sulfonic acid groups.

If you're interested I can send you a reprint
of a 2001 paper in Biotech. Histochem. on dye
nomenclature; just let me know where to send
it to. It's similar to Ch. 3 in Conn's Biol. St.

Conn's 10th edn uses US spellings (despite
a Brit publisher) but in dye names this differs 
only slightly from English spelling. The only
differences I can think of offhand are having
an f instead of ph in parts of words
derived from sulfur, and e rather than ae in
hematoxylin and hematein. 

We hope suppliers will adopt Conn's as a
standard for names - not only spellings
but also calling the dyes what they really
are. Ethyl green is an example of one
that's probably been wrongly labeled for
30+ years despite the fact that it's
better for its job than methyl green.

Best regards,   John.
________________________________________
Mark Ray wrote:
> John,
> 
> Help me out here.   Is the terminal e required in safranin_e_?  It's an
> amine, right?  I've noticed that JT Baker has added the e on its MSD
> Sheet but not in its catalog, as yet.   Are the spellings in the new
> Conn to be considered universally authoritative or at least  correct in
> US dialect?  I think I may have a lot of catalog, product label, and
> MSD Sheet revision to do.
> 
> Regards,
> Mark Ray
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
> 
> John Kiernan wrote:
> 
> >Point taken! I notice that Chroma do use
> >the correct English spelling of fuchsine.
> >Like nearly everyone else they spell
> >phloxin with a terminal e, which is wrong
> >because it's in the same group as eosin.
> >Chroma also put a wrong terminal e on their
> >English spelling of erythrosin; that one is
> >unusual.
> >              --- John Kiernan
> >------------------------------------
> >Gudrun Lang wrote:
> >
> >
> >>Dear John,
> >>I think the German spelling is Fuchsin (without e). Please look at
> >>www.chroma.de at their catalogue.
> >>Your vixen is German "Fchsin" (fuechsin).
> >>
> >>greetings from Austria
> >>Gudrun
> >>
> >>----- Original Message -----
> >>From: "John Kiernan" <jkiernan <@t> uwo.ca>
> >>To: "Rena" <RFail <@t> Charleston.net>; <histonet <@t> lists.utsouthwestern.edu>
> >>Sent: Thursday, September 30, 2004 5:29 AM
> >>Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels]
> >>
> >>Dear Rena Fail,
> >>
> >>Red alkaline phosphatase product.
> >>
> >>Methods giving permanently mountable red products have been available
> >>for many years.
> >>Davis & Ornstein (1959) introduced hexazotized pararosaniline as a
> >>trifunctional trapping agent for naphthols released by hydrolysis of
> >>naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of
> >>the four components of basic fuchsine and is commercially available in
> >>fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is
> >>closely similar to pararosaniline and also available in pure form
> >>(Horobin, 2002); It is currently preferred for making the hexazotized
> >>reagent.
> >>
> >>Kits are sold, but the substrate solution is easily made in the lab. In
> >>a simple procedure published at the IHCWorld.com (2004) web site (see
> >>below for its URL), the working mixture is prepared from four stock
> >>solutions, which are stored at 4 C. Solution D is warmed to room
> >>temperature before using. The amount of levamisole (inhibitor of
> >>endogenous tissue alkaline phosphatase) is clearly stated.
> >>
> >>The working solution is made immediately before using. Mix 10 drops each
> >>of A and B, then add 10 drops of solution C followed by 10 Kits are
> >>sold, but the substrate solution is easily made in the lab. In a simple
> >>procedure published at the IHCWorld.com (2004) web site, the working
> >>mixture is prepared from four stock solutions, which are stored at 4 C.
> >>Solution D is warmed to room temperature before using. I've annotated
> >>the instructions from IHCworld.com to simplify the weighing and
> >>measuring.
> >>
> >>A.  New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M)
> >>B.  Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole
> >>hydrochloride) 0.48% in water.
> >>C.  Naphthol AS-BI phosphate, 1% in 100% dimethylformamide.
> >>D.  0.05 M Tris-HCl buffer, pH 8.7.
> >>
> >>The working solution is made immediately before using. Mix 10 drops each
> >>of A and B, then add 10 drops of solution C followed by 10ml of the
> >>buffer (D). A drop is assumed to be 0.05 ml.
> >>
> >>Sections are incubated in the working solution for 10-20 minutes at room
> >>temperature. They are then washed counterstained as desired, dehydrated,
> >>cleared and coverslipped.
> >>
> >>Lojda et al (1979, p. 74-75) warn that the working solution may be red
> >>or brown from colored impurities in new fuchsine or pararosaniline. Such
> >>solutions cause excessive yellow background staining. The yellow/brown
> >>impurities are the same ones that can make Schiff's reagent yellow
> >>(removable from Schiff with activated charcoal). Pararosaniline
> >>certified by the Biological Stain Commission is suitable for making
> >>alkaline phosphatase substrate mixtures because it is required to be
> >>substantially free of brown and yellow materials (Penney et al., 2002).
> >>
> >>Probably any batch of basic fuchsine certified by the Biological Stain
> >>Commission will be OK for making the IHCWorld.com alkaline phosphatase
> >>substrate solution, because each of the four possible components of
> >>basic fuchsine has three diazotizable amino groups and their molecular
> >>weights are all quite close. I don't know of any published study that
> >>establishes this, so you're probably safest to go with either certified
> >>pararosaniline or with new fuchsine from a reputable dealer.
> >>
> >>References.  (Please check them out if you can. The 2nd is easy enough!)
> >>
> >>Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298.
> >>
> >>IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution.
> >>http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm
> >>
> >>Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed.
> >>Oxford:BIOS.
> >>
> >>Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer.
> >>
> >>Penney et al. (2002) Biotech. Histochem. 77:237-275.
> >>
> >>Finally, please note that the correct spelling is fuchsine, not fuchsin,
> >>despite anything you might read in a catalog or on a bottle label. This
> >>is not a matter of American, British and German spellings. (Fuchsin is
> >>German for vixen; the dyes are, I think, named from their colours being
> >>similar to those of Fuchsia flowers rather than foxes.) Look in
> >>Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an
> >>informal name of a chemical indicates that it is a base, often an amine.
> >>The -in ending occurs with compounds that are not bases, such as
> >>dextrin, eosin etc.
> >>
> >>                  John Kiernan
> >>                  London, Canada
> >>------------------------------------------------------------------
> >>Rena wrote:
> >>
> >>
> >>>I have been having some trouble with some Abs stained with New Fuchsin
> >>>and permanent red. Recently I was asked if I were using Levamisole in
> >>>the permanent red and if so how much? I was told too much Levamisole in
> >>>permanent red would result in no staining. I have used 1 drop per ml for
> >>>both New Fuchsin and permanent red, which is the appropriate amount.  We
> >>>have run both DABS and either permanent  red or New Fuchsin with the
> >>>same AB from the same vial with vastly different results. Any ideas?
> >>>Rena Fail
> >>>_______________________________________________
> >>>Histonet mailing list
> >>>Histonet <@t> lists.utsouthwestern.edu
> >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>>
> >>>
> >>_______________________________________________
> >>Histonet mailing list
> >>Histonet <@t> lists.utsouthwestern.edu
> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>
> >>_______________________________________________
> >>Histonet mailing list
> >>Histonet <@t> lists.utsouthwestern.edu
> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>
> >>
> >
> >
> >

Mark Ray wrote:
> 
> John Kiernan wrote:
> 
> >Point taken! I notice that Chroma do use
> >the correct English spelling of fuchsine.
> >Like nearly everyone else they spell
> >phloxin with a terminal e, which is wrong
> >because it's in the same group as eosin.
> >Chroma also put a wrong terminal e on their
> >English spelling of erythrosin; that one is
> >unusual.
> >              --- John Kiernan
> >------------------------------------
> >Gudrun Lang wrote:
> >
> >
> >>Dear John,
> >>I think the German spelling is Fuchsin (without e). Please look at
> >>www.chroma.de at their catalogue.
> >>Your vixen is German "Füchsin" (fuechsin).
> >>
> >>greetings from Austria
> >>Gudrun
> >>
> >>----- Original Message -----
> >>From: "John Kiernan" <jkiernan <@t> uwo.ca>
> >>To: "Rena" <RFail <@t> Charleston.net>; <histonet <@t> lists.utsouthwestern.edu>
> >>Sent: Thursday, September 30, 2004 5:29 AM
> >>Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels]
> >>
> >>Dear Rena Fail,
> >>
> >>Red alkaline phosphatase product.
> >>
> >>Methods giving permanently mountable red products have been available
> >>for many years.
> >>Davis & Ornstein (1959) introduced hexazotized pararosaniline as a
> >>trifunctional trapping agent for naphthols released by hydrolysis of
> >>naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of
> >>the four components of basic fuchsine and is commercially available in
> >>fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is
> >>closely similar to pararosaniline and also available in pure form
> >>(Horobin, 2002); It is currently preferred for making the hexazotized
> >>reagent.
> >>
> >>Kits are sold, but the substrate solution is easily made in the lab. In
> >>a simple procedure published at the IHCWorld.com (2004) web site (see
> >>below for its URL), the working mixture is prepared from four stock
> >>solutions, which are stored at 4 C. Solution D is warmed to room
> >>temperature before using. The amount of levamisole (inhibitor of
> >>endogenous tissue alkaline phosphatase) is clearly stated.
> >>
> >>The working solution is made immediately before using. Mix 10 drops each
> >>of A and B, then add 10 drops of solution C followed by 10 Kits are
> >>sold, but the substrate solution is easily made in the lab. In a simple
> >>procedure published at the IHCWorld.com (2004) web site, the working
> >>mixture is prepared from four stock solutions, which are stored at 4 C.
> >>Solution D is warmed to room temperature before using. I've annotated
> >>the instructions from IHCworld.com to simplify the weighing and
> >>measuring.
> >>
> >>A.  New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M)
> >>B.  Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole
> >>hydrochloride) 0.48% in water.
> >>C.  Naphthol AS-BI phosphate, 1% in 100% dimethylformamide.
> >>D.  0.05 M Tris-HCl buffer, pH 8.7.
> >>
> >>The working solution is made immediately before using. Mix 10 drops each
> >>of A and B, then add 10 drops of solution C followed by 10ml of the
> >>buffer (D). A drop is assumed to be 0.05 ml.
> >>
> >>Sections are incubated in the working solution for 10-20 minutes at room
> >>temperature. They are then washed counterstained as desired, dehydrated,
> >>cleared and coverslipped.
> >>
> >>Lojda et al (1979, p. 74-75) warn that the working solution may be red
> >>or brown from colored impurities in new fuchsine or pararosaniline. Such
> >>solutions cause excessive yellow background staining. The yellow/brown
> >>impurities are the same ones that can make Schiff's reagent yellow
> >>(removable from Schiff with activated charcoal). Pararosaniline
> >>certified by the Biological Stain Commission is suitable for making
> >>alkaline phosphatase substrate mixtures because it is required to be
> >>substantially free of brown and yellow materials (Penney et al., 2002).
> >>
> >>Probably any batch of basic fuchsine certified by the Biological Stain
> >>Commission will be OK for making the IHCWorld.com alkaline phosphatase
> >>substrate solution, because each of the four possible components of
> >>basic fuchsine has three diazotizable amino groups and their molecular
> >>weights are all quite close. I don't know of any published study that
> >>establishes this, so you're probably safest to go with either certified
> >>pararosaniline or with new fuchsine from a reputable dealer.
> >>
> >>References.  (Please check them out if you can. The 2nd is easy enough!)
> >>
> >>Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298.
> >>
> >>IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution.
> >>http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm
> >>
> >>Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed.
> >>Oxford:BIOS.
> >>
> >>Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer.
> >>
> >>Penney et al. (2002) Biotech. Histochem. 77:237-275.
> >>
> >>Finally, please note that the correct spelling is fuchsine, not fuchsin,
> >>despite anything you might read in a catalog or on a bottle label. This
> >>is not a matter of American, British and German spellings. (Fuchsin is
> >>German for vixen; the dyes are, I think, named from their colours being
> >>similar to those of Fuchsia flowers rather than foxes.) Look in
> >>Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an
> >>informal name of a chemical indicates that it is a base, often an amine.
> >>The -in ending occurs with compounds that are not bases, such as
> >>dextrin, eosin etc.
> >>
> >>                  John Kiernan
> >>                  London, Canada
> >>------------------------------------------------------------------
> >>Rena wrote:
> >>
> >>
> >>>I have been having some trouble with some Abs stained with New Fuchsin
> >>>and permanent red. Recently I was asked if I were using Levamisole in
> >>>the permanent red and if so how much? I was told too much Levamisole in
> >>>permanent red would result in no staining. I have used 1 drop per ml for
> >>>both New Fuchsin and permanent red, which is the appropriate amount.  We
> >>>have run both DABS and either permanent  red or New Fuchsin with the
> >>>same AB from the same vial with vastly different results. Any ideas?
> >>>Rena Fail
> >>>_______________________________________________
> >>>Histonet mailing list
> >>>Histonet <@t> lists.utsouthwestern.edu
> >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>>
> >>>
> >>_______________________________________________
> >>Histonet mailing list
> >>Histonet <@t> lists.utsouthwestern.edu
> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>
> >>_______________________________________________
> >>Histonet mailing list
> >>Histonet <@t> lists.utsouthwestern.edu
> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>
> >>
> >
> >
> >