49026 Milmont Drive, Fremont, CA 94538

Tel : +1 (800) 421-4149, Fax: +1 (510) 824-1490,

support@biogenex.com

Emergo Europe, Molenstraat 15, NL-2513 BH The Hague, The Netherlands

Tel: (+31) 70-345-8570, Fax: (+31) 70-346-7299

SPECIAL STAINS

Van Gieson Staining

Cat. No. SS044-YAK
Doc. No. 932-SS044-YAK
Release Date: 12-Feb-2014

Rev. No. A

Intended Use
This product is intended for in vitro diagnostic use on the i6000 BioGenex Automated Staining System & also for Manual
staining.
Summary and Explanation
Van Gieson Stain is used to differentiate between collagen and smooth muscle in tumors and to demonstrate the increase of
collagen in diseases. This method combines two or more anionic dyes and rely on differential binding by tissue components.
The differentiation is determined by a combination of differences in the relative size of the dye molecules, differences in the
physical structure of the tissue, and differences in the amino acid composition of tissue Elements.
Principles of the Procedure
Van Gieson's Stain is a mixture of Picric Acid and Acid Fuchsin. It is the simplest method of differential staining
of Collagen and other Connective Tissue. When using combined solutions of picric acid and acid fuchsin, the small molecules of
picric acid penetrate all of the tissues rapidly, but are only firmly retained in the close textured, red blood cells and muscle. The
larger molecules of Ponceau S displace picric acid molecules from collagen fibres, which have larger pores, and allow the larger
molecules to enter.
Reagents Provided
Celestin blue Solution (HX931-YAK)
Mayers Haematoxylin (HX790-YAK)
Curtis Stain (HX937-YAK)

1 x 20 mL, ready to use

1 x 20 mL, ready to use
1 x 20 mL, ready to use

Reagents Required but Not Supplied

EZ-DeWax Solution (HK584-5K)

1 x 500 ml, concentrate

Super SensitiveTM Wash Buffer, 20X (HK583-5K)

1 x 500 ml, concentrate

Mounting Medium
Positive Control Tissue
Storage and Handling
Store all reagents at 2-8C. Do not use after expiration dates as indicated on the reagent labels
Specimen Preparation
Fixation plays an important role in preserving the tissue structures to be visualized using this stain. 10% neutral buffered
formalin is the preferred fixative, however, other fixatives like Carnoys may also be used. Ensure that the fixed sections are
adequately embedded in paraffin. Cut tissue sections to 4-5 microns.

49026 Milmont Drive, Fremont, CA 94538

Tel : +1 (800) 421-4149, Fax: +1 (510) 824-1490,
support@biogenex.com

Emergo Europe, Molenstraat 15, NL-2513 BH The Hague, The Netherlands

Tel: (+31) 70-345-8570, Fax: (+31) 70-346-7299

Precautions
Not regarded as a health or environmental hazard under current legislation. This product contains no hazardous material at a
reportable concentration according to U.S.29 CFR 1910.1200 and EC Directive 91/155/EC.
BioGenex i6000
Automated Staining System Protocol
1.
2.
3.

4.

Bring reagents to room temperature. Deparaffinize the tissues and hydrate to distilled water.
Load the barcode-labeled slides for the appropriate stain into the BioGenex Automated Staining System slide racks. Load
the barcode-labeled reagent vials into the BioGenex Automated Staining System reagent racks.
Start Special Stains software on the instrument and check the protocol parameters against the factory default settings listed
below. Change any of the parameters as required. Please note that any parameter change, once saved, becomes the default
setting until changed again. Use of parameter settings other than factory default requires validation by the user.
Select Start Scan and the instrument will perform the steps listed in the table (if the factory default settings have been
selected).

Van Gieson Staining Protocol Default Setting:

Reagent
Special Stains Wash Solution

No. of Incubations
3

Incubation Time
30 sec.

No. of After Rinses

0

Celestin Blue

5 min

Haematoxylin

5 min

Curtis stain

10 min

3*

* Alcohol Wash
Mounting Instructions
After staining, dehydrate and clear in xylene as usual and apply permanent mounting medium.
Manual Protocol for Alcian Blue PAS Stain
1.
2.
3.
4.
5.
6.

Deparaffinize and hydrate the slides to distilled water.

Apply 250l Celestin blue solution for 5 min.
Rinse in distilled water.
Add 250l Haematoxylin solution and let stand for 5 minutes. Wash well in running tap water.
Add 250l Curtis stain, allow to stand for 10 min.
Blot. Dehydrate rapidly in alcohols, clear and mount

Quality Control
A positive control slide one that will display positive staining with this stain should be included in every run.
Trouble shooting
1. Follow data sheet instructions correctly.
2. Gently invert all reagents prior to use.
Expected Results
Nuclei stain blue; Collagen stain bright red; Cytoplasm, muscle, fibrin and red blood cells stain yellow
Limitations of the procedure
1. The solution weakens after long standing and may be strengthened by adding few drops of fresh Acid Fuchsin.
2. The thickness of the section may affect the intensity of the staining. Necrotic tissue may exhibit non-specific staining.
3. Use tissue, which shows positive structure or element for which it is being tested.

49026 Milmont Drive, Fremont, CA 94538

Tel : +1 (800) 421-4149, Fax: +1 (510) 824-1490,
support@biogenex.com

Emergo Europe, Molenstraat 15, NL-2513 BH The Hague, The Netherlands

Tel: (+31) 70-345-8570, Fax: (+31) 70-346-7299

Performance Characteristics
Specificity of Special Stain
Van Gieson is used to differentiate between collagen and smooth muscle in tumors and to demonstrate the increase of collagen
in diseases.
BioGenex has conducted studies to evaluate the performance of all its Special Stains products. BioGenex Special Stains have
shown reproducible and consistent results when used within a single run, between runs and between lots. The products have
been determined to be stable for the periods specified on the labels either by standard real time or accelerated testing methods.
BioGenex ensures product quality through 100% quality control for all products released and through surveillance programs.
Bibliography
1. Manual of Histopathological and Histochemical Procedures, 16th ed. Compiled by JP Pickett, Pathology Dept., Duke
Medical Center, Durham, NC, 1983.
2. Theory and Practice of Histotechnological Techniques, 4th ed., JD Bancroft & A Stevens, eds., Churchill Livingstone, New
York(NY), 1996.
3. Handbook of Histopathological and Histochemical Techniques Ed.3, Butterworth, London, UK