Periodic Acid-Schiff (PAS With Diastase) Cat. No. SS039-25D

49026 Milmont Drive, Fremont, CA 94538 Emergo Europe, Molenstraat 15, NL-2513 BH The Hague, The Netherlands
Tel : +1 (800) 421-4149, Fax: +1 (510) 824-1490,

support@biogenex.com
SPECIAL STAINS
Periodic Acid-Schiff (PAS; with Diastase)
Cat. No. SS039-25D
Doc. No. 932-SS039-25D Rev. No. B
Release Date: 09-Nov-2015
Intended Use
This product is intended for in vitro diagnostic use on the BioGenex Xmatrx® Automated Staining System.
Summary and Explanation
The PAS reaction in tissue sections is useful for outlining tissue structures such as basement membranes, capsules and blood
vessels. This staining procedure may also be used for the demonstration of fungal organisms in tissue sections. The diastase
(Alpha-amylase) digestion procedure, followed by staining with PAS, is useful as an aid in the evaluation of glycogen storage
disease.
Principles of the Procedure

Glycols are oxidized to aldehydes when reacted with periodic acid. The glycol-containing cellular components are stained upon
addition of Schiff’s reagent, a mixture of pararosaniline and sodium metabisulfite, which causes a pararosaniline adduct to be released.
Polysaccharides present in the tissue are hydrolyzed to starch, glycogen and degradation products from Diastase digestion.
Reagents Provided
Periodic Acid Solution (HX890-07D) 1 x 7 ml ready to use
Schiff’s reagent (HX702-07D) 1 x 7ml ready to use
Scott’s Solution (HX725-07D) 1 x 7ml ready to use
Hematoxylin Solution (HX790-07D) 1 x 7ml ready to use
Diastase Kit for diastase digestion:

Alpha Amylase, HK718-5K
Diluent for Amylase, HX719-5X
Reagents Required but Not Supplied

EZ-DeWax™ Solution (HK584-5K) 1 x 500 ml, concentrate
TM
Super Sensitive Wash Buffer, 20X (HK583-5K) 1 x 500 ml, concentrate
Mounting Medium
Positive Control Tissue
Storage and Handling
Store all reagents at 2-8°C. 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 or Bouin's Solution may be used. Ensure that the fixed sections are adequately embedded in paraffin. Cut tissue
sections to 4-5 microns.
1
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Precautions
F, R11, Xi, R36, R67 = highly flammable, Irritating to eyes. Vapours may cause drowsiness and dizziness.
S26, S38, S45, S53, S24/25, S36/37/39, S60, P11 = In case of contact with eyes, rinse immediately with plenty of water and seek
medical advice. In case of insufficient ventilation, wear suitable respiratory equipment. In case of accident or if you feel unwell, seek
medical advice immediately (show the label where possible). Avoid exposure - obtain special instructions before use. Avoid contact
with skin and eyes. Wear suitable protective clothing, gloves and eye/face protection. This material and its container must be
disposed of as hazardous waste.
Preparation of Working Solutions

Prepare Diastase Working Solution by pouring a part of the Diastase Diluent into one vial of Diastase powder. Invert several times to
mix. Pour the mixed solution into the Diastase Diluent vial for use on the instrument. Diastase working Solution is stable for 1 week
when stored at 2-8°C. Bring to room temperature before use .
BioGenex XmatrxAutomated Staining System Protocol

1. Bring reagents to room temperature.
2. Load the barcode-labeled slides for the appropriate stain into the Xmatrx Automated Staining System slide racks. Load the
RFID Tagged vials into the Xmatrx Automated Staining System reagent racks.
3. 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.
4. Select “Start Scan” and the instrument will perform the steps listed in the table (if the factory default settings have been selected).
Mounting Instructions
After staining, wipe slides, air dry, then dehydrate in 100% alcohol. Clear in xylene and apply permanent mounting medium.
PAS PROTOCOL DEFAULT SETTING:
Reagent No. of Incubations Incubation Time No. of After Rinses

Baking 1 15 min. 0
EZ-DeWax™ 3 3 min. 0
Special Stains Wash Solution 3 20 sec. 0
Diastase Working Solution 2* 30 min. 3
Periodic Acid Solution 1 10 min. 2
Schiff's Reagent 1 15 min. 1
o
Scotts Solution 1 5 min at 40 C 2
Hematoxylin Solution 1 3 min. 4
Clearmount/Alcohol Wash 1 1 min. 0
XMount 1 10 min 0
* For Pas with Diastase digestion only.
Quality Control
A positive control slide—one that will display positive staining with this stain—should be included in every run. This product has
been validated through extensive testing on the liver tissue.
2
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Trouble shooting
1. Follow data sheet instructions correctly
2. Make sure all reagents are mixed properly prior to use, to prevent any settling down of components
3. Large tissue sections require more regents. Depending on the size of the tissue set the parameters correctly
4. Mount slides as per data sheet
Expected Results
Nuclei stain blue; glycogen and other carbohydrates oxidized to aldehyde by periodate stain red to pink. There is no stained glycogen
seen in diastase-digested tissue.
Limitations of the procedure

1. The thickness of the section may affect the intensity of the staining. Necrotic tissue may exhibit non-specific staining.
2. All tissues need to be dewaxed. If a different type of wax is used, the staining may not penetrate into the tissue.
3. Do not leave the slides overnight without mounting, this dries up the dye. Mount slides as soon as the run is over.
4. Use tissue, which shows positive structure or element for which it is being tested
Performance Characteristics
Specificity of Special Stain
The PAS stains specifically glycogen, glycoproteins, mucopolysaccharides, basement membrane and mucin.
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
nd
1. Sheehan DC & Hrapchak BB: Theory and Practice of Histotechnology, 2 ed. CV Mosby, St. Louis, (MO), 52 & 164-167, 1980.
th
2. Culling CFA, et al: Cellular Pathology Technique, 4 ed. Butterworths, 216-220, 1985.
3. Hotchkiss RD: A microchemical reaction resulting in the staining of polysaccharide structures. Arch Biochem 16:131, 1948.
nd
4. Davey FR, Nelson DA: Periodic Acid Schiff (PAS) Stain, Hematology, 2 ed. WJ Williams, et al, eds. McGraw-Hill, New
York,1630-1632, 1977.
5. Thompson SW: Selected Histochemical and Histopathological Methods, CC Thomas, Springfield, (IL), 520-539, 1966.
th
6. Theory and Practice of Histotechnological Techniques, 4 ed., JD Bancroft & A Stevens, eds., Churchill Livingstone, New York
(NY), 1996.

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49026 Milmont Drive, Fremont, CA 94538 Emergo Europe, Molenstraat 15, NL-2513 BH The Hague, The Netherlands

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

Summary and Explanation

Principles of the Procedure

Diastase Kit for diastase digestion:

Reagents Required but Not Supplied

Storage and Handling

Preparation of Working Solutions

BioGenex XmatrxAutomated Staining System Protocol

PAS PROTOCOL DEFAULT SETTING:

Reagent No. of Incubations Incubation Time No. of After Rinses

Limitations of the procedure

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