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FETAL HEMOGLOBIN-DIFFERENTIAL STAIN KIT
(MODIFIED KLEIHAUER TECHNIQUE)

(UNAVAILABLE)

Solution I - Citric Acid 0.lM
Solution II - Disodium Phosphate 0.2M
Solution III - Mayer’s Acid Alum Hematoxylin
Solution IV - 0.1% Aqueous Eosin

This procedure, a modification of the Kleihauer Technique, is based on the resistance of fetal hemoglobin to acid conditions. By drying and fixation with ethanol, hemoglobin is precipitated within the red cells. Air-dried and fixed blood films are treated with a citric acid-phosphate buffer of low pH. Adult hemoglobin is rapidly eluted but HbF remains precipitated and is stained with eosin.

RECOMMENDED PROCEDURE:   Prepare thin (monolayer) blood films of capillary or fresh EDTA-anticoagulated blood and fix in 80% ethanol for 5 minutes. Rinse with water and allow to dry. Pipette 37.7 ml of Sol. I and 12.3 ml of Sol. II into a coplin jar and place in a 37oC water bath. When solution reaches 37oC, place fixed slide in coplin jar for 5 minutes. Move slides up and down a few times to ensure adequate elution of hemoglobin from cells. Remove slides and rinse with water. Stain with Sol. III for 1 - 4 minutes. Rinse with water. Stain with Sol. IV for 1 minute. Rinse with water and dry. Examine under low power for occasional eosin staining cells. Oil immersion is used for the pale ghosts cells that do not stain with eosin.

RESULTS:  RBC’s containing HbF stain brilliantly with eosin, while those without HbF appear as colorless "ghosts". The hematoxylin stains the WBC nuclei to prevent misinterpretation as eosin-positive cells.

# 5700

Fetal Hemoglobin-Differential Stain Kit
1 Kit (250 ml Ea. Sol.)

N/A
# 5702

Sol. I - Citric Acid 0.lM
1 Pack (Two 250 ml Btls.)

N/A
# 5703

Sol. I - Citric Acid 0.lM
1 Btl. (250 ml)

N/A
# 5706

Sol. II - Disodium Phosphate 0.2M
1 Pack (Two 250 ml Btls.)

N/A
# 5707 Sol. II - Disodium Phosphate 0.2M
1 Btl. (250 ml)
N/A
# 5710 Sol. III - Mayer’s Acid Alum Hematoxylin
1 Pack (Two 250 ml Btls.)
N/A
# 5711 Sol. III - Mayer’s Acid Alum Hematoxylin
1 Btl. (250 ml)
N/A
# 5714 Sol. IV - 0.1% Aqueous Eosin
1 Pack (Two 250 ml Btls.)
N/A
# 5715 Sol. IV - 0.1% Aqueous Eosin
1 Btl. (250 ml)
N/A

CONTENTS:  Citric Acid, Disodium Phosphate, Hematoxylin, Sodium Iodate, Potassium Alum, Eosin

These solutions are made from certified dyes (when applicable).
FOR IN VITRO DIAGNOSTIC USE ONLY.

FETAL HEMOGLOBIN - DIFFERENTIAL STAINING KIT
(MODIFIED KLEIHAUER OR MODIFIED CLAYTON TECHNIQUE)
ROOM TEMPERATURE PROCEDURE (RTP)

(UNAVAILABLE)

Modified Kleihauer Technique:
Solution I - Fixative
Solution II - Citric Acid/Phosphate Buffer
Solution III - Mayer’s Acid Alum Hematoxylin
Solution IV - Eosin 0.1% Aqueous

Modified Clayton Technique:
Solution I - Fixative
Solution II - Citric Acid /Phosphate Buffer
Solution III - Eosin 0.1% Aqueous

This product has been developed to employ a new modified Kleihauer procedure for fetal hemoglobin that does not require heat as in the old modified Kleihauer technique. This test can now be performed at room temperature. We recommend the use of hematoxylin for staining the WBC’s so as to avoid misidentification of eosin-positive cells. This is done with the new Kleihauer technique rather than the new Clayton technique.

These procedures are based upon the resistance of fetal hemoglobin to acid conditions. Fixation of the smear allows hemoglobin to precipitate within the RBC. The Citric acid/Phosphate buffer solution eludes the adult hemoglobin. Fetal hemoglobin remains precipitated and stains bright red with eosin, making for easy differentiation. In the new and old Kleihauer procedure, WBC’s are counterstained with hematoxylin to avoid misidentification as eosin-positive cells.

RECOMMENDED PROCEDURE:  Prepare thin (monolayer) blood films of capillary or fresh EDTA-anticoagulated blood and fix in Sol. I for 5 minutes. Rinse with distilled water and allow to dry. Place slide in Sol.II or flood to elude 5 - 10 minutes. Rinse with distilled water. Stain with Sol. III for 1 - 4 minutes and rinse with distilled water. Stain with Sol. IV for 2 - 6 minutes. Rinse again in distilled water and allow time to dry.

RESULTS:  RBC’s containing HbF stain brilliantly with eosin, while those without HbF appear as colorless "ghosts". The hematoxylin stains the nuclei of the WBC’s blue to prevent misinterpretation as eosin-positive cells.

# 5720

Fetal Hemoglobin - Differential Stain Kit
(RTP) (Modified Kleihauer Technique)

1 Kit (120 ml Ea. Sol.)

N/A

# 5721

Fetal Hemoglobin - Differential StainKit
(RTP) (Modified Clayton Technique)

1 Kit (120 ml Ea. Sol.)

N/A
# 5722

Solution I - Fixative
1 Pack (Two 120 ml Btls.)

N/A
# 5723

Solution I - Fixative
1 Btl. (120 ml)

N/A
# 5726

Solution II - Citric Acid/Phosphate Buffer
1 Pack (Two 120 ml Btls.)

N/A
# 5727

Solution II - Citric Acid/Phosphate Buffer
1 Btl. (120 ml)

N/A
# 5730

Solution III - Mayer’s Acid Alum Hematoxylin
1 Pack (Two 120 ml Btls.)

N/A
# 5731

Solution III - Mayer’s Acid Alum Hematoxylin
1 Btl. (120 ml)

N/A
# 5734

Solution IV - 0.1% Aqueous Eosin (Kleihauer)
1 Pack (Two 120 ml Btls.)

N/A
# 5735

Solution IV - 0.1% Aqueous Eosin (Kleihauer)
1 Btl. (120 ml)

N/A
# 5736

Solution III - 0.1% Aqueous Eosin (Clayton)
1 Pack (Two 120 ml Btls.)

N/A
# 5737

Solution III - 0.1% Aqueous Eosin (Clayton)
1 Btl. (120 ml)

N/A

CONTENTS:  Ethanol, Citric Acid, Disodium Phosphate, Hematoxylin, Sodium Iodate, Potassium Alum, Eosin.

These solutions are made from certified dyes (when applicable).
FOR IN VITRO DIAGNOSTIC USE ONLY.

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