[Histonet] Re: Marking inks

Tony Henwood AnthonyH <@t> chw.edu.au
Mon Oct 4 17:57:36 CDT 2010

The following might be of use

Marking surgical margins are often required for determining the adequacy
of excision of lesions. Sometimes up to 6 margins (medial, lateral,
superior, inferior, superial and deep margins) are required to be
individually marked.

The properties of a good marker include:

1.	It is quick to dry. 
2.	It is easy to apply. 
3.	It has a long shelf life. 
4.	It is cheap. 
5.	It is non-toxic. 
6.	It does not spread beyond the edge of the marked area. 
7.	It is preserved during processing (no contamination of
8.	It is visible macroscopically and microscopically. 
9.	Sectioning is not affected. 
10.	It is radiolucent. 
11.	There are multiple colours available.

India Ink, a colloidal suspension of particulate carbon in aqueous gum
has been used for many years, but is messy, slow to dry and spreads
readily over the tissue surface. 10% Silver Nitrate in methanol has been
suggested but it also spreads over the surface and is macroscopically

5% alcian blue in 80% ethanol is an excellent tissue marker. It is easy
to apply, quick to dry, has a long shelf life and is preserved during

Birch et al (1990) have suggested that specimens can be "dunked" into 1%
aqueous alcian blue. This method is quick, cheap and the coating is

Harris (1990) has suggested the use of Tippex fluid. This is a solution
of titanium dioxide and polyacrylate in trichloethane. It has a long
shelf life, is easy to use, quick drying, non-toxic and the tissue
processor is not affected. The marker does however have a tendency to
lift and is radiodense.

Hunter-Craig et al (1991) rolled blotted dry specimens in starch powder.
The surface coating of starch is visible on light microscopy but
strikingly apparent if crossed polarisers are used.

Artist's pigments in acetone (a 50% solution) were used by Patterson and
Davies (1988). The pigments are quick drying, have a good range of
different colours that are visible both macroscopically and
microscopically (especially using polarised light) and are resistant to
processing. They are however, radiodense and have a short storage life.
It must be remembered that the dry powders (which contain cobalt,
manganese and cadmium) are toxic. The pigment granules are also of
similar size and density to microcalcification and there may be
confusion on subsequent specimen mammography after tissue slicing.

Suggested pigments are as follows:

Cobalt Blue	Blue	Blue
Alizarin Crimson	Red	Red 
Viridian	Green	Green

Armstrong et al (1990) have described the use of organically coloured
gelatines, where 8% solution of dye is prepared in 24% aqueous gelatine.
They found it easier to discriminate between colours of particulate dyes
(i.e. plant substances), with the gelatine staining with both
haematoxylin and eosin giving a pink to purple colour. They found clear
demarcation between two adjacent colours and only a modest knowledge of
botany was required for the identification of the three plant materials.

Janus Green	Blue	Purple 
India Ink	Black	Black  
Paprika	Red/brown	Red-pigmented cellulose 
Tumeric 	Yellow	Cerebriform starch granules
Henna	Brown	Brown-pigmented cellulose
Bismark Brown	Brown	Brown particles

Unfortunately tumeric causes knife scratching.

Armstrong et al (1990) J.Clin.Pathol., 43:604-607
Birch et al (1990) J.Clin.Pathol 43:608-609
Harris (1990) J.Clin.Pathol 43:346
Hunter-Craig et al (1991) J.Clin.Pathol. 44:874-875
Patterson & Davies (1988) J.Clin.Pathol 41:1013-1016


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Robert
Sent: Saturday, 2 October 2010 3:48 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Marking inks

Richard Cartun asks about marking inks used for surgical pathology

if I just need one color (great majority of specimens) I use india ink
bought from a stationer or craft supply store - cheap, easily replaced,
and it gets the job done.

If I need colors, I prefer the Davidson marking inks, available through
some though not all vendors. They offer seven colors, including orange
(they have to sell their product in Tennessee and Texas, after all!) and
purple. (I have no commercial connection with Davidson inks.)

I blot my specimens thoroughly dry, and don't use a fixative. If I did,
I'd use white vinegar (5% acetic acid), possibly diluted 1:1 with water.
Bouin's fixative and acetone both have obvious environmental problems.

Some people use tattoo inks. Cheap and available in a huge number of
colors, but you have to read some truly disgusting catalogs to get them.

I didn't know about green Davidson inks gumming up processors - thanks
for that tip. (I wonder why other inks should be different.)

With all marking inks, it's very important to re-cap the bottle as soon
as you're through with it. If you re-cap promptly, an opened bottle will
keep for years.

Bob Richmond
Samurai Pathologist
Knoxville TN

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