Re: [Histonet] Stain vs dye and control

[Rene J Buesa]

In a very simplistic way, a "dye" is the active chemical substance (usually an 
aniline powder) that you use to prepare a staining solution for certain "stain" 
procedure or method."Dye" is a more "precise" term but stain had "many shades 
of color"!
René J.  

 On Friday, March 6, 2015 3:49 PM, Jorge A. Santiago-Blay 
 wrote:
   

 Dear Histonetters:

Last semester I taught a microbiology lab and, as I was reviewing for
class, noticed some lack of precision in the use of the terms "dye" vs.
"stain" in biology. Could someone help?

While I am on "stains", I have been following the emails on controls and
wonder a couple of things.

1. What would testing for bacteria on beef jerky, hot dogs or burgers
accomplish that is different (ideally better) that what one accomplishes by
pulling out from fresh cultures out of a medium (e.g. liquid, such as
broth, solid, such as slabs, agar)?  Is it the idea to test for bacteria in
an animal tissue? If so, would a solid medium (like someone mentioned
recently, such as agar) do?

2. An advantage of using fresh bacterial cultures of known Gram is that it
could be used to test whether the reagents are good enough. Last semester I
had the suspicion that one (or more) of our Gram reagents where not up to
par.

If you have any feedback, please feel free to email me directly at
blayjo...@gmail.com . Thank you.

Sincerely,

Jorge

Jorge A. Santiago-Blay, PhD
blaypublishers.com
http://blayjorge.wordpress.com/
http://paleobiology.si.edu/staff/individuals/santiagoblay.cfm
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[Histonet] Stain vs dye and control

[Jorge A. Santiago-Blay]

Dear Histonetters:

Last semester I taught a microbiology lab and, as I was reviewing for
class, noticed some lack of precision in the use of the terms "dye" vs.
"stain" in biology. Could someone help?

While I am on "stains", I have been following the emails on controls and
wonder a couple of things.

1. What would testing for bacteria on beef jerky, hot dogs or burgers
accomplish that is different (ideally better) that what one accomplishes by
pulling out from fresh cultures out of a medium (e.g. liquid, such as
broth, solid, such as slabs, agar)?  Is it the idea to test for bacteria in
an animal tissue? If so, would a solid medium (like someone mentioned
recently, such as agar) do?

2. An advantage of using fresh bacterial cultures of known Gram is that it
could be used to test whether the reagents are good enough. Last semester I
had the suspicion that one (or more) of our Gram reagents where not up to
par.

If you have any feedback, please feel free to email me directly at
blayjo...@gmail.com . Thank you.

Sincerely,

Jorge

Jorge A. Santiago-Blay, PhD
blaypublishers.com
http://blayjorge.wordpress.com/
http://paleobiology.si.edu/staff/individuals/santiagoblay.cfm
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[Histonet] RE: Mushrooms for GMS fungus control

[Norton, Sally]

We have used orange rind and brie - left to grow mold  - as controls in a 
pinch.  Our docs preferred the cheese.

Sally Norton
Seattle Children's Hospital

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Pence
Sent: Friday, March 06, 2015 9:38 AM
To: 'Piche, Jessica'; Jeffrey Robinson; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Mushrooms for GMS fungus control

As your fungal control?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Piche, Jessica
Sent: Friday, March 06, 2015 11:32 AM
To: Jeffrey Robinson; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Mushrooms for GMS fungus control

I will say I hacked a mushroom off of a tree once and processed it and it 
stained nicely for GMS. I also used some chicken that was in the fridge too 
long and we actually use that for our GMS control!!

Jessica Piche, HT(ASCP)
Waterbury Hospital

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jeffrey Robinson
Sent: Friday, March 06, 2015 12:16 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Mushrooms for GMS fungus control

How about mushrooms?  Has anyone had any success using mushrooms as a GMS 
fungus control?

Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA


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[Histonet] RE: Mushrooms for GMS fungus control

[Mike Pence]

As your fungal control?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Piche, Jessica
Sent: Friday, March 06, 2015 11:32 AM
To: Jeffrey Robinson; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Mushrooms for GMS fungus control

I will say I hacked a mushroom off of a tree once and processed it and it 
stained nicely for GMS. I also used some chicken that was in the fridge too 
long and we actually use that for our GMS control!!

Jessica Piche, HT(ASCP)
Waterbury Hospital

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jeffrey Robinson
Sent: Friday, March 06, 2015 12:16 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Mushrooms for GMS fungus control

How about mushrooms?  Has anyone had any success using mushrooms as a GMS 
fungus control?

Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA


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and is not intended for any unauthorized person. If you, the reader, are not 
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[Histonet] RE: Mushrooms for GMS fungus control

[Piche, Jessica]

I will say I hacked a mushroom off of a tree once and processed it and it 
stained nicely for GMS. I also used some chicken that was in the fridge too 
long and we actually use that for our GMS control!!

Jessica Piche, HT(ASCP)
Waterbury Hospital

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jeffrey Robinson
Sent: Friday, March 06, 2015 12:16 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Mushrooms for GMS fungus control

How about mushrooms?  Has anyone had any success using mushrooms as a GMS 
fungus control?

Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA


This email and attachments may contain PHI that is privileged and confidential 
and is not intended for any unauthorized person. If you, the reader, are not 
the intended recipient you are hereby notified that any dissemination, 
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[Histonet] Re: CONTROLS

[Deann D. Chandler]

I once tried using Slim Jims as a control for gram staining.  It had a lot of 
positive bacteria, but no negatives.  We then went to our microbiology lab got 
a little colony of positive and negative bacteria, embedded it in agar and 
processed it.  This worked great!  I do want to see if this works with 
hamburger or hot dogs.

DeAnn Chandler, BS, HT (ASCP)
Histology Coordinator
Newberry County Memorial Hospital

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[Histonet] Mushrooms for GMS fungus control

[Jeffrey Robinson]

How about mushrooms?  Has anyone had any success using mushrooms as a GMS 
fungus control?

Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA


This email and attachments may contain PHI that is privileged and confidential 
and is not intended for any unauthorized person. If you, the reader, are not 
the intended recipient you are hereby notified that any dissemination, 
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Re: AW: [Histonet] Trichrome & Fixation

[John Kiernan]

Quite right! Treating sections of formaldehyde-fixed tissue with Bouin's fluid 
is not mordanting; binding site retrieval is probably a more accurate term. 

Picric acid induces a change in the section that improves the selective uptake 
of the heteropolyacid and the larger dye anions by collagen and of the smaller 
dye anions by cytoplasm. Picric acid alone is just as good as Bouin for this 
purpose. The picric acid treatment has the effect of making a section of 
formaldehyde-fixed tissue more like a section of tissue fixed in a picric 
acid-containing fixative such as Bouin or Gendre. For spectacular trichrome 
staining you need a fixative that contains mercuric chloride, like Zenker or 
SuSa.

An example of a mordant in biological staining is iron alum in Heidenhain's 
method. The subsequently applied haematein solution forms a coloured complex 
with the iron bound by the sections. The differentiation in iron alum solution 
then removes bound haematein by forming a water-soluble complex. The word 
"mordant" is probably best avoided for staining with pre-formed dye-metal 
complexes such as carmine, aluminium-haematein (haemalum), Weigert's 
iron-haematein and gallocyanine-chrome alum. See R.W.Horobin (1982) 
"Histochemistry" Stuttgart: Gustav Fischer (ISBN 3437107003 or 0407002480), 
page 77.

John Kiernan
= = =
On 06/03/15, Gudrun Lang   wrote:
> I disagree. Usually a dye binds covalently to the mordant, and a mordant 
> should be a multi-valent metal (like aluminium, ferrum, molybden,..). The 
> dye-mordant-complex binds via the mordant to the substrate.
> Picric acid of Bouin's is washed out before staining.
> 
> It may be a matter of definition of the term mordant.
> 
> Gudrun
> 
> -Ursprüngliche Nachricht-
> Von: Johnson, Carole [mailto:cjohn...@nmda.nmsu.edu] 
>  
> Gesendet: Freitag, 06. März 2015 15:10
> An: gu.l...@gmx.at
> Betreff: RE: [Histonet] Trichrome & Fixation
> 
> Boiuns solution acts as a mordant in trichrome stains
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] 
>  On Behalf Of Gudrun Lang
> Sent: Friday, March 06, 2015 1:13 AM
> To: 'Amos Brooks'
> Cc: histonet@lists.utsouthwestern.edu
> Subject: AW: [Histonet] Trichrome & Fixation
> 
> Hi,
> years ago we did a stain called CAB (=one step trichrome) regularly on 
> liver-tissue. I don't know if it was because of ignorance or with aim, but it 
> was done without Bouin. The result was blue-grey hepatocytes and darker blue 
> collagen. - also totally different to the result with Bouin (red hepatocytes).
> 
> I think the Bouin is less a "re-fixation" than more an "binding-site 
> retrival" in this context.
> 
> Gudrun
> 
> 
> -Ursprüngliche Nachricht-
> Von: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] 
>  Im Auftrag von Amos Brooks
> Gesendet: Donnerstag, 05. März 2015 22:01
> An: histonet@lists.utsouthwestern.edu
> Betreff: [Histonet] Trichrome & Fixation
> 
> Hi,
>  It is interesting that you should mention the importance of fixation on the 
> Trichrome stain. I have an image of two murine hearts processed, cut and 
> stained side by side. The only difference between the two is that they were 
> harvested at different times, so one sat in formalin long enough to be 
> properly fixed the other one was placed in the fixative then immediately 
> brought in to be processed from 70% ethanol on. They are *totally* different 
> looking. The red muscle tissue looks more purple therefore less distinct from 
> the blue blood vessels. You get a similar effect with lung bronchial 
> epithelium.
> 
> Cheers,
> Amos
> 
> On Tue, Mar 3, 2015 at 1:00 PM, 
> wrote:
> 
> > Message: 15
> > Date: Tue, 03 Mar 2015 12:34:33 -0500
> > From: John Kiernan 
> > Subject: Re: [Histonet] Masson trichrome and H and E
> > To: Emily Brown ,
> > "histonet@lists.utsouthwestern.edu"
> > 
> > Message-ID: <7390afa3112a.54f5a...@uwo.ca>
> > Content-Type: text/plain; CHARSET=US-ASCII
> >
> > If you can't get two colours with H&E, don't expect to get the colour 
> > scheme right with Masson's trichrome, which needs more skill. If you 
> > are hoping to show basement membranes in the kidney, you would do 
> > better to use a technically simpler staining method such as 
> > picro-sirius red or periodic acid-Schiff. If for some reason you 
> > really need three colours, a one-step trichrome such as Gomori's, 
> > Cason's or Gabe's might be the way to go rather than Masson's or one 
> > of the other multi-step trichromes. Remember that all trichrome 
> > methods are greatly influenced by the fixative. A post-fixation 
> > treatment of the sections, usually with picric acid, is needed for 
> > formaldehyde-fixed tissues. Some alternative post-fixation treatments 
> > were proposed by Yu & Chapman (2003) J. Histotechnol. 26(2): 131-134, but 
> > their coloured photos were not very convincing.
> >
> > Making up staining soluti

[Histonet] Richard Allan Tissue Marking Dye or Cancer Diagnostics

[Vickroy, James]

In the past I have always used Cancer Diagnostics marking dyes.   Can someone 
comment on the Richard Allan dyes?  Do they work as well as the Cancer 
diagnostic ones?   Does the dye remain well after processing so that it is 
easily seen in the sections?

Jim

Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvick...@springfieldclinic.com


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AW: [Histonet] Trichrome & Fixation

[Gudrun Lang]

I disagree. Usually a dye binds covalently to the mordant, and a mordant should 
be a multi-valent metal (like aluminium, ferrum, molybden,..). The 
dye-mordant-complex binds via the mordant to the substrate.
Picric acid of Bouin's is washed out before staining.

It may be a matter of definition of the term mordant.

Gudrun

-Ursprüngliche Nachricht-
Von: Johnson, Carole [mailto:cjohn...@nmda.nmsu.edu] 
Gesendet: Freitag, 06. März 2015 15:10
An: gu.l...@gmx.at
Betreff: RE: [Histonet] Trichrome & Fixation

Boiuns solution acts as a mordant in trichrome stains

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang
Sent: Friday, March 06, 2015 1:13 AM
To: 'Amos Brooks'
Cc: histonet@lists.utsouthwestern.edu
Subject: AW: [Histonet] Trichrome & Fixation

Hi,
years ago we did a stain called CAB (=one step trichrome) regularly on 
liver-tissue. I don't know if it was because of ignorance or with aim, but it 
was done without Bouin. The result was blue-grey hepatocytes and darker blue 
collagen.  - also totally different to the result with Bouin (red hepatocytes).

I think the Bouin is less a "re-fixation" than more an "binding-site retrival" 
in this context.

Gudrun


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Amos Brooks
Gesendet: Donnerstag, 05. März 2015 22:01
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Trichrome & Fixation

Hi,
 It is interesting that you should mention the importance of fixation on 
the Trichrome stain. I have an image of two murine hearts processed, cut and 
stained side by side. The only difference between the two is that they were 
harvested at different times, so one sat in formalin long enough to be properly 
fixed the other one was placed in the fixative then immediately brought in to 
be processed from 70% ethanol on. They are *totally* different looking. The red 
muscle tissue looks more purple therefore less distinct from the blue blood 
vessels. You get a similar effect with lung bronchial epithelium.

Cheers,
Amos

On Tue, Mar 3, 2015 at 1:00 PM, 
wrote:

> Message: 15
> Date: Tue, 03 Mar 2015 12:34:33 -0500
> From: John Kiernan 
> Subject: Re: [Histonet] Masson trichrome and H and E
> To: Emily Brown ,
> "histonet@lists.utsouthwestern.edu"
> 
> Message-ID: <7390afa3112a.54f5a...@uwo.ca>
> Content-Type: text/plain; CHARSET=US-ASCII
>
> If you can't get two colours with H&E, don't expect to get the colour 
> scheme right with Masson's trichrome, which needs more skill. If you 
> are hoping to show basement membranes in the kidney, you would do 
> better to use a technically simpler staining method such as 
> picro-sirius red or periodic acid-Schiff. If for some reason you 
> really need three colours, a one-step trichrome such as Gomori's, 
> Cason's or Gabe's might be the way to go rather than Masson's or one 
> of the other multi-step trichromes. Remember that all trichrome 
> methods are greatly influenced by the fixative. A post-fixation 
> treatment of the sections, usually with picric acid, is needed for 
> formaldehyde-fixed tissues. Some alternative post-fixation treatments 
> were proposed by Yu & Chapman (2003) J. Histotechnol. 26(2): 131-134, but 
> their coloured photos were not very convincing.
>
> Making up staining solutions in-house is always cheaper than buying 
> pre-made solutions.
>
> John Kiernan
> London, Canada
>
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AW: [Histonet] Trichrome & Fixation

[Gudrun Lang]

Hi,
years ago we did a stain called CAB (=one step trichrome) regularly on 
liver-tissue. I don't know if it was because of ignorance or with aim, but it 
was done without Bouin. The result was blue-grey hepatocytes and darker blue 
collagen.  - also totally different to the result with Bouin (red hepatocytes).

I think the Bouin is less a "re-fixation" than more an "binding-site retrival" 
in this context.

Gudrun


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Amos Brooks
Gesendet: Donnerstag, 05. März 2015 22:01
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Trichrome & Fixation

Hi,
 It is interesting that you should mention the importance of fixation on 
the Trichrome stain. I have an image of two murine hearts processed, cut and 
stained side by side. The only difference between the two is that they were 
harvested at different times, so one sat in formalin long enough to be properly 
fixed the other one was placed in the fixative then immediately brought in to 
be processed from 70% ethanol on. They are *totally* different looking. The red 
muscle tissue looks more purple therefore less distinct from the blue blood 
vessels. You get a similar effect with lung bronchial epithelium.

Cheers,
Amos

On Tue, Mar 3, 2015 at 1:00 PM, 
wrote:

> Message: 15
> Date: Tue, 03 Mar 2015 12:34:33 -0500
> From: John Kiernan 
> Subject: Re: [Histonet] Masson trichrome and H and E
> To: Emily Brown ,
> "histonet@lists.utsouthwestern.edu"
> 
> Message-ID: <7390afa3112a.54f5a...@uwo.ca>
> Content-Type: text/plain; CHARSET=US-ASCII
>
> If you can't get two colours with H&E, don't expect to get the colour 
> scheme right with Masson's trichrome, which needs more skill. If you 
> are hoping to show basement membranes in the kidney, you would do 
> better to use a technically simpler staining method such as 
> picro-sirius red or periodic acid-Schiff. If for some reason you 
> really need three colours, a one-step trichrome such as Gomori's, 
> Cason's or Gabe's might be the way to go rather than Masson's or one 
> of the other multi-step trichromes. Remember that all trichrome 
> methods are greatly influenced by the fixative. A post-fixation 
> treatment of the sections, usually with picric acid, is needed for 
> formaldehyde-fixed tissues. Some alternative post-fixation treatments 
> were proposed by Yu & Chapman (2003) J. Histotechnol. 26(2): 131-134, but 
> their coloured photos were not very convincing.
>
> Making up staining solutions in-house is always cheaper than buying 
> pre-made solutions.
>
> John Kiernan
> London, Canada
>
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