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Product Information May-Grünwald Giemsa
Intended Use 3. Prepare the Giemsa working solution:
May-Grünwald’s eosin methylene blue and Giemsa’s azure Dilute 50 ml Giemsa solution with 450 ml Sørensen
eosin methylene blue are intended to be used for staining of buffer solution pH 7.
blood and bone marrow smears and cytological specimens, 4. Proceed according to the table below
such as urine sediment or sputum. For staining of most Reagent sequence Minutes
histology specimen (mostly gastric sections) Giemsa is used. Methanol or undiluted May-Grünwald 3
Principle May-Grünwald working solution 5
The purple colour of cell nuclei, is due to molecular Move slides gently in Sørensen buffer pH 7 1
interaction between eosin Y and an azure B-DNA complex. Move slides gently in Giemsa working 20
The intensity of the staining depends on the azure B content solution
and on the ratio azure B/eosin Y. The pH of the solutions, Flush in tap or de-ionized water
fixation and staining time can influence the staining result. Note: When using methanol in the first step as fixative,
basophilic granules might be missed. This is known from
Composition literature.
May-Grünwald is alcohol based and contains
May-Grünwald’s eosin methylene blue and methanol (>85%). C. Traditional method according to Pappenheim for whole
Giemsa is alcohol based and contains Giemsa’s azure eosin blood smears
methylene blue, methanol (>50%) and glycerol. 1. Prepare Sørensen buffer solution pH 7:
Dilute 100 ml of the concentrated Sørensen phosphate
Stability and Storage buffer with 1.9L de-ionized water.
May-Grünwald and Giemsa are stable for five years. The 2. Prepare the Giemsa working solution:
bottles must be kept closed. The advised storage temperature Dilute 25 ml Giemsa solution with 475 ml Sørensen
is 18 - 30°C. Used solutions and solutions that are past their buffer solution pH 7.
shelf-life must be disposed of, according to local disposal 3. Proceed according to the table below
guidelines. Reagent sequence Minutes
Procedures for bone marrow, cytology samples and Undiluted May-Grünwald 3
blood smears Flush in de-ionised water 1
A. General method for Bone marrow or Cytology specimen: Move slides gently in Giemsa working 20
1. Prepare Sørensen buffer solution pH 7: solution
Dilute 100 ml of the concentrated Sørensen phosphate Flush in tap or de-ionised water
buffer with 1.9L de-ionized water.
2. Prepare the May-Grünwald working solution: D. Quick staining method for whole blood smears
Dilute 250 ml May-Grünwald solution with 250 ml 1. P repare Sørensen buffer solution pH 7:
Sørensen buffer solution pH 7. Dilute 100 ml of the concentrated Sørensen phosphate
3. Prepare the Giemsa working solution: buffer with 1.9L de-ionized water.
Dilute 50 ml Giemsa solution with 450 ml Sørensen 2. P repare the Giemsa working solution:
buffer solution pH 7. Dilute Giemsa solution 1 to 6 up to 1 to 8 with Sørensen
4. Proceed according to the table below buffer solution pH 7.
Reagent sequence Minutes 3. Proceed according to the table below
Undiluted May-Grünwald 3 Reagent sequence Minutes
May-Grünwald working solution 5 Undiluted May-Grünwald 2-3
Move slides gently in Sørensen buffer pH 7 1 Move slides gently in Sørensen buffer pH 7 1
Move slides gently in Giemsa working 25 Move slides gently in Giemsa working 4-5
solution solution
Flush in tap or de-ionized water Flush in tap or de-ionised water
B. G eneral method for whole blood smears: E. Staining method for whole blood smears using a bridge
1. Prepare Sørensen buffer solution pH 7: 1. P repare Sørensen buffer solution pH 7:
Dilute 100 ml of the concentrated Sørensen phosphate Dilute 100 ml of the concentrated Sørensen phosphate
buffer with 1.9L deionized water. buffer with 1.9L de-ionized water.
2. Prepare the May-Grünwald working solution: 2. P repare the Giemsa working solution:
Dilute 250 ml May-Grünwald solution with 250 ml Dilute Giemsa solution 1 to 1 up to 1 to 4 with Sørensen
Sørensen buffer solution pH 7. buffer solution pH 7.
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3. Proceed according to the table below power when heavily used and the staining times should hloe
Reagent sequence Minutes be longer or fresh solutions should be used. SC
Undiluted May-Grünwald 2-3
Flush in tap or de-ionized water 1 melet
Giemsa working solution 4-5
Flush in tap or de-ionized water
Note: The times as listed in the tables are ndray
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approximates and can be adjusted to suit personal m
preferences. Staining solutions will lose their staining
Performance Characteristics
Type Blood-cell Amount Characteristics kohden
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RBC 4 – 6 x 10 /liter Pink/brown discs; clearer in the middle due to their concave structure hon
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PLT 0.2 – 0.3 x 10 /liter Purple coloured granules; much smaller then RBC
NEUT 50 – 70 % * Transparent, pink/blue cytoplasm; 2-5 lobed bright purple nucleus
EO 2 – 4 % Typical pink-orange granulated cytoplasm; generally 2-lobed purple hee
nucleus P
LYM 20 – 40 % Transparent purple cytoplasm; one large, purple-pink nucleus or
MONO 3 – 8 % Largest of the leukocytes; transparent, pink/blue cytoplasm with
horseshoe-shaped pink/purple nucleus
BASO 0.5 – 1.0 % Granulo-rich cytoplasm exhibiting dark-blue stain overruling the C
dark-blue nucleus stain ea
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* From total WBC population; generally 5 – 7 x 10 cells/liter
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s may-grünwald stained blood cells (100x) I
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troubleshooting bone marrow, Cytology samples and blood smears U
Insufficient colour development of leucocytes
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- Prepared smears not completely dried Colour intensity bone marrow, Cytology samples and
- Glass slides used are not degreased or pre-treated blood smears ontrol
- Exhausted May-Grünwald or Giemsa working solution Varying times may influence the colour intensity of both the C
Too much erythrocyte fragments blue and red:
- Specimen older than 12 hours Step Step Blue Red
- Blood smear dried at ambient temperature > 30°C May-Grünwald Giemsa S
- Prepared smear not completely dried + 5 min. + 5 min. ++ ++
Colour too much blue and red leaner
- Too long staining in May-Grünwald and Giemsa + 5 min. same time + C
- Too short colour development in Sørensen buffer or same time + 5 min. +
de-ionized water – 5 min. − 5 min. −− −−
- Ambient temperature > 30°C
– 5 min. same time – S
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Procedure for histology samples Colour intensity histology samples
1. Prepare the Giemsa working solution: Varying staining times may influence the colour intensity of
Add 20 ml Giemsa solution to 80 ml de-ionized water. both the blue and red:
It is important to add the Giemsa to the water and not Step Step Blue Red
vice versa. May-Grünwald Giemsa
Filter the solution before use. + 5 min. + 5 min. ++ ++
2. Prepare the differentiation solution:
Add 4 drops of Glacial Acetic Acid (96%) to 100 ml of + 5 min. same time +
de-ionized water. same time + 5 min. +
Measure the pH of the solution. It should be – 5 min. − 5 min. −− −−
3.0 – 3.2.
3. Proceed according to the tables below – 5 min. same time –
same time − 5 min. −
t de-paraffination of tissue
Reagent sequence Minutes Frequently asked Questions
UltraClear/Xylene 3x1 Do I have to change my working protocol?
Ethanol 100% 3x1 When changing to J.T.Baker there’s no real need to change
Ethanol 70% 1 the whole procedure. For an optimized performance we
Flush with tap water 1 refer to the Product Information.
See scheme above.
t Staining of tissue Are the solutions ready to use?
Reagent sequence Minutes This depends on the application. We refer to the methods
Insert 3 times in de-ionized water advised. In general May-Grünwald is used undiluted or
Giemsa working solution (use only once) 30 diluted 1 to 1. Giemsa is always diluted in the range from
dip just 1:5 up to 1:20.
Differentiation fluid (differentiation to purple) once For which specimen types is May-Grüwald and Giemsa
Ethanol 96% (differentiation to blue) dip just suitable?
once Blood smears, bone marrow and tissue material, such as
Isopropanol (2-propanol) dip just gastric sections.
once Features and benefits
Isopropanol (2-propanol) 3 x 2
UltraClear/Xylene (refresh each time) 3 x 2 Optimized formulation Colours are balanced
Coverslip slides
Optimized working Colour tones easy adaptable
Performance Characteristics for histology samples procedure available
Nucleus : blue / violet
Cytoplasm : blue Sørensen buffer solution Easy diluting (if needed),
Erythrocytes : pink 3716 available reproducible results
Eosinophilic granules : orange
Basophilic granules : purple Traditional production Avoids crystallization
method
troubleshooting for histology samples
Insufficient colour development Suitable for hematology, Broadly applicable
- Glass slides used are not degreased or pre-treated histology and cytology
- Exhausted Giemsa working solution
Colour too much blue
- Too long staining in Giemsa
- Ambient temperature > 30°C
- Slide too short in differentiation fluid
Colour too much red instead of purple
- Too much glacial Acetic Acid in differentiation fluid
- Slide to long inserted in differentiation fluid
Poor colour development of blue
- Too long inserted in Ethanol 96%
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ordering Information hloe
Product Product Number Packaging SC
May-Grünwald 3855.0500 500 ml
May-Grünwald 3855.1000 1 liter melet
May-Grünwald 3855.2500 2.5 liter
Giemsa 3856.0500 500 ml ndray
Giemsa 3856.1000 1 liter I
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Giemsa 3856.2500 2.5 liter
Sørensen buffer concentrate 3716 10 x 100 ml
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