Sigma-Aldrich 1.00485.0001 Manual

1.00485.0001
Microscopy
Masson-Goldner staining kit
for the visualization of connective tissue with
trichromic staining
For professional use only
In Vitro Diagnostic Medical Device
Intended purpose
This "Masson-Goldner staining kit - for the visualization of connective tis-
sue with trichromic staining" is used for human-medical cell diagnosis and
serves the purpose of the histological investigation of sample material of
human origin.
The Masson-Goldner staining method is a trichrome stain that is primarily
used for imaging connective-tissue structures in organs. It is a ready-to-use
staining kit that when used together with other in vitro diagnostic products
from our portfolio makes target structures evaluable for diagnostic purposes
(by fixing, embedding, staining, counterstaining, mounting) in histological
specimen materials, for example histological sections of e. g. the liver, the
kidney, the intestine, the placenta and similar.
Unstained structures are relatively low in contrast and are extremely diffi-
cult to distinguish under the light microscope. The images created using the
staining solutions help the authorized and qualified investigator to better
define the form and structure in such cases. Further examinations may be
necessary to reach a definitive diagnosis.
Principle
Using a combination of three different staining solutions, muscle fibers, col-
lagenous fibers, fibrin and erythrocytes can be selectively visualized.
The original methods were primarily used to differentiate collagenous and
muscle fibers. The stains used have different molecular sizes and enable the
individual tissues to be stained differentially.
The Masson-Goldner staining technique can be carried out using formalin
fixed material. Subsequent to staining the nucleus with Weigert's iron hema-
toxylin, components such as muscle, cytoplasm and erythrocytes are stained
with azophloxin and orange G solution. Connective tissue is then counter
stained using light green SF solution.
Sample material
Starting materials are sections of tissue embedded in paraffin (3 - 5 µm
thick paraffin sections).
Reagents
Cat. No. 1.00485.0001
Masson-Goldner staining kit
for the visualization of connective tissue with trichromic staining
Package components:
The staining kit contains
Reagent 1: Azophloxine solution
Reagent 2: Tungstophosphoric acid orange G solution
Reagent 3: Light green SF solution
Reagent 4: Acetic acid 10%
Also required:
Cat. No. 1.15973 Weigert's iron hematoxylin kit
for nuclear staining in histology
Sample preparation
The sampling must be performed by qualified personnel.
All samples must be treated using state-of-the-art technology.
All samples must be clearly labeled.
Suitable instruments must be used for taking samples and their prepara-
tion. Follow the manufacturer's instructions for application / use.
When using the corresponding auxiliary reagents, the corresponding in-
structions for use must be observed.
Deparaffinize and rehydrate sections in the conventional manner.
Reagent preparation
The reagents of the Masson-Goldner staining kit - for the visualization of
connective tissue with trichromic staining are ready-to-use, dilution of the
solutions is not necessary and merely produces a deterioration of the stain-
ing result and their stability.
Acetic acid 1%
For preparation of approx. 100 ml solution mix:
Reagent 4 (Acetic acid 10%)
Distilled water
Weigert's iron hematoxylin staining solution
Mix Weigert's solution A and Weigert's solution B (from Cat. No. 1.15973)
in the ratio 1 + 1.
The prepared staining solution remains stable for approx. one working
week.
The solution must be exchanged as soon as the cell nuclei have been
stained brown.
Procedure
Staining in the staining cell
Deparaffinize histological slides in the conventional manner and rehydrate
in a descending alcohol series.
The stated times should be adhered to in order to guarantee an optimal
staining result.
Slide with histological specimen
Weigert's iron hematoxylin staining solution
Running tap water
Acetic acid 1%*
Reagent 1 (Azophloxine solution)
Acetic acid 1%*
Reagent 2 (Tungstophosphoric acid orange G solution)
Acetic acid 1%*
Reagent 3 (Light green SF solution)
Acetic acid 1%*
Ethanol 70%
Ethanol 96%
Ethanol 100%
Ethanol 100%
Ethanol 100%
Xylene or Neo-Clear
Xylene or Neo-Clear
Mount the Neo-Clear
slides with e. g. Entellan
* Prepare fresh acetic acid 1% regularly.
500 ml
500 ml
500 ml
Note: If the expected result of the staining of muscle fibers is too weak or
500 ml
not present at all, the specimen should be incubated in Bouin's so-
lution at 37°C for 30 min before the staining procedure and after
rehydration. The specimen must then be rinsed in distilled water. The
staining procedure starts with Weigert's iron hematoxylin staining
2x 500 ml
solution (see "Procedure").
After dehydration (ascending alcohol series) and clarification with xylene or
Neo-Clear , histological slides can be covered with non-aqueous mounting
TM
agents (e. g. Entellan
be stored.
Result
Nuclei
Cytoplasm, muscle
Connective tissue, acidic mucous substances
Erythrocytes
www.sigmaaldrich.com
TM
TM
-wet slides with Neo-Mount
TM TM
new and cover glass.
TM
new, Neo-Mount ) and a cover glass and can then
TM TM
dark brown to black
brick red
green
bright orange
EN
10 ml
90 ml
5 min
5 min
approx. 30 sec
10 min
approx. 30 sec
1 min
approx. 30 sec
2 min
approx. 30 sec
30 sec
30 sec
30 sec
30 sec
2 min
5 min
5 min
or the xylene-wet