1.09468.

0100
1.11373.0100 Reagents
1.11374.0100 Cat. No. 109468

Buffer tablets pH 7.2
for preparing buffer solution acc. to WEISE
100 tabs

for staining of blood smears

Microscopy
Cat. No. 111373 Buffer tablets pH 6.4 100 tabs
for preparing buffer solution acc. to WEISE
Buffer tablets pH 7.2 for the staining of blood smears

Cat. No. 111374 Buffer tablets pH 6.8 100 tabs

for preparing buffer solution acc. to WEISE for preparing buffer solution acc. to WEISE
for staining of blood smears for the staining of blood smears

Also required:
Buffer tablets pH 6.4 for Giemsa stain:
Cat. No. 106009 Methanol for analysis EMSURE® 1 l, 2.5 l, 5 l
for preparing buffer solution acc. to WEISE ACS,ISO,Reag. Ph Eur
for the staining of blood smears Cat. No. 109204 Giemsa’s azur eosin methylene blue 100 ml, 500 ml,
solution 1 l, 2.5 l
for microscopy
Buffer tablets pH 6.8 for May-Grünwald stain:
Cat. No. 101424 May-Grünwald’s eosine-methylene blue 100 ml, 500 ml,
for preparing buffer solution acc. to WEISE solution modified 1 l, 2.5 l
for staining of blood smears for microscopy
for Wright stain:
Cat. No. 101383 Wright’s eosin methylene blue solution 100 ml, 500 ml,
for microscopy 2.5 l
For professional use only
for Leishman stain:
Cat. No. 105387 Leishman’s eosin methylene blue solution 500 ml
modified
In Vitro Diagnostic Medical Device for microscopy

Intended purpose
These
Buffer tablets pH 6.4 - for preparing buffer solution acc. to WEISE for the
staining of blood smears
Buffer tablets pH 6.8 - for preparing buffer solution acc. to WEISE for the
staining of blood smears
Buffer tablets pH 7.2 - for preparing buffer solution acc. to WEISE for stain-
ing of blood smears
are used to prepare buffer solutions according to WEISE and pH-buffered
staining solutions. These solutions are used in conjunction with other in
vitro diagnostic products from our portfolio for the reproducible staining of
sample materials of human origin, serving to make target structures evalu-
able for diagnostic purposes (by fixing, embedding, staining, counterstain-
ing, mounting) in haematological and clinico-cytological specimen material,
such as whole blood and bone-marrow smears.
The type of buffer tablet that is appropriate for use from case to case is de-
scribed in the corresponding instructions for use for our staining solutions,
solid dyes, and test kits.
Phosphate buffer solutions acc. to WEISE are used for Giemsa’s, May-
Grünwald’s, combination of Giemsa’s/May-Grünwald’s (“Pappenheim stain”),
Wright’s, Leishman’s and Hemacolor® stainings (Cat. No.111661 and
111674) are used as solutions for rinsing and for dilution if necessary.
Principle
Neutral distilled water is required for blood smear staining to obtain repro-
ducible staining results. It is necessary both for preparing dilute Giemsa’s
solution and for rinsing stained slides.
Freshly distilled water can be stabilized by adding a buffer. The buffer mix-
ture acc. to WEISE is an effective means for this purpose. Upon addition of
this buffer mixture the pH is automatically adjusted to the required value.
Fluctuations in the pH of distilled water are caused on the one hand by
atmospheric carbon dioxide and on the other by detergent residues on
equipment. Acidic distilled water produces a red-tinted stain, and alkaline
distilled water a blue-tinted stain.
Buffered solutions of pH 6.4, 6.8, or 7.2 acc. to WEISE reliably prevent
discolorations.
Buffer solution acc. to WEISE pH 6.4 shows a result with reddish
stained erythrocytes and red-violet stained cells with nuclei,
buffer solution acc. to WEISE pH 6.8 shows reddish stained erythro-
cytes and more intense red-violet stained cells with nuclei,
buffer solution acc. to WEISE pH 7.2 shows reddish-brownish stained
erythrocytes and even more intensive red-violet stained cells with nuclei.
Sample preparation
The sampling must be performed by qualified personnel.
Please use thin, air-dried blood or bone-marrow smears, as well as cytologi-
cal materials, that have been stored not longer than three days.
The smears must be dried in air for at least 30 minutes and be fixed ac-
cording to the relevant instructions prior to the actual reaction.
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.
Reagent preparation
Buffer solution acc. to WEISE pH 6.4, 6.8, or 7.2
For preparation of approx. 1000 ml of solution, add and dissolve:
Buffer tablet
Cat. No. 111373 (pH 6.4) or
Cat. No. 111374 (pH 6.8) or 1
Cat. No. 109468 (pH 7.2)
depending on the required reaction color
Distilled water 1000 ml
The buffer solution can be used for at least 4 weeks when kept in a tightly
closed glass bottle.
Dilute Giemsa’s staining solution for manual staining
For preparation of approx. 200 ml solution mix:
Giemsa’s azur eosin methylene blue solution 10 ml
Buffer solution acc. to WEISE pH 6.4, 6.8, or 7.2 190 ml
Mix well, leave to stand for 10 min, and filter if necessary
Procedure
The stated times should be adhered to in order to guarantee an optimal
staining result.
Giemsa stain on the staining rack

Slide with air-dried smear

Sample material
Only fresh, native blood or bone-marrow smears should be used as the start- Methanol 3 min
ing material for all stains.
Dilute Giemsa’s staining solution for manual staining 20 min
Further, also clinical cytological material can be used: urine sediment,
sputum, fine needle aspiration biopsies (FNAB), imprints, rinses, effusions, Buffer solution acc. to WEISE 1 min
exsudates.
Buffer solution acc. to WEISE 1 min
Air-dry (e. g. over night or at 50 °C in the drying cabinet)
May-Grünwald stain on the staining rack Buffer solution acc. to WEISE pH 6.8

Slide with air-dried smear Giemsa May-Grün- Wright Leishman

stain wald stain stain
May-Grünwald’s eosine- 1 ml cover completely 3 min stain
methylene blue solution
Nuclei resp. red to violet red-violet red-violet red-violet
modified chromatin
Buffer solution acc. to 1 ml mix 6 min Cytoplasm of blue blue blue blue
WEISE lymphocytes
Buffer solution acc. to rinse 1 min Cytoplasm of grey-blue grey-blue grey-blue grey-blue
WEISE monocytes

Air-dry (e. g. over night or at 50°C in the drying cabinet) neutrophilic light violet light violet light violet light violet
granula

Wright stain on the staining rack eosinophilic reddish to red- brick-red brick-red to brick-red
granula brown red-brown
Slide with air-dried smear basophilic dark violet dark violet to dark violet to dark violet
granula black black
Wright’s eosin methylene 1 ml cover completely 1 min
blue solution Thrombocytes violet violet violet violet

Buffer solution acc. to 1 ml mix 4 min Erythrocytes reddish reddish reddish reddish
WEISE
Buffer solution acc. to rinse 1 min Buffer solution acc. to WEISE pH 7.2
WEISE
Giemsa May-Grün- Wright Leishman
Buffer solution acc. to rinse 1 min stain wald stain stain
WEISE stain

Air-dry (e. g. over night or at 50°C in the drying cabinet) Nuclei resp. violet violet blue-violet red-violet
chromatin

Cytoplasm of blue blue blue blue

Leishman stain on the staining rack lymphocytes

Slide with air-dried smear Cytoplasm of grey-blue grey-blue grey-blue grey-blue

monocytes
Leishman’s eosin methylene 1 ml cover completely 1 min
blue solution modified neutrophilic light violet light violet light violet light violet
granula
Buffer solution acc. to 2 ml mix 5 min
eosinophilic reddish to red-brown red to red- brick-red
WEISE
granula red-brown brown
Buffer solution acc. to rinse 1 min basophilic dark violet dark violet to dark violet to dark violet
WEISE granula black black
Buffer solution acc. to rinse 1 min Thrombocytes violet violet violet violet
WEISE
Erythrocytes reddish-brow- pink to brow- pink to brow- reddish
Air-dry (e. g. over night or at 50°C in the drying cabinet) nish nish nish

To enable hematology specimens to be stored over a period of several

months, it is advisable to cover them with a mounting medium (e. g.
Neo-Mount®, DPX new, Entellan® new) and a cover glass. The stained prep-
arations had to be very well dried. When left unmounted, the stain remains
stable for about 3 days, covered with immersion oil only for a few hours.
The use of immersion oil is recommended for the analysis of stained slides
with a microscopic magnification >40x.
Technical notes
The microscope used should meet the requirements of a medical diagnostic
laboratory.
When using histoprocessors and automatic staining systems, please follow
the instructions for use supplied by the supplier of the system and software.
Remove surplus immersion oil before filing.

Diagnostics
Result
Diagnoses are to be made only by authorized and qualified personnel.
Buffer solution acc. to WEISE pH 6.4
Valid nomenclatures must be used.
This method can be supplementarily used in human diagnostics.
Giemsa May-Grün- Wright Leishman
stain wald stain stain Further tests must be selected and implemented according to recognized
stain methods.
Suitable controls should be conducted with each application in order to
Nuclei resp. red to violet red-violet red-violet red-violet avoid an incorrect result.
chromatin

Cytoplasm of blue blue blue blue Storage

lymphocytes
Store the
Cytoplasm of grey-blue grey-blue grey-blue grey-blue Buffer tablets pH 6.4 - for preparing buffer solution acc. to WEISE for the
monocytes staining of blood smears
neutrophilic light violet light violet light violet light violet
Buffer tablets pH 6.8 - for preparing buffer solution acc. to WEISE for the
granula staining of blood smears
Buffer tablets pH 7.2 - for preparing buffer solution acc. to WEISE for
eosinophilic reddish to red- brick-red to brick-red to brick-red staining of blood smears
granula brown red-brown red-brown at +5 °C to +30 °C.
basophilic dark violet dark violet to dark violet to dark violet
granula black black Shelf-life
Thrombocytes violet violet violet violet The Buffer tablets pH 6.4 - for preparing buffer solution acc. to WEISE for
the staining of blood smears
Erythrocytes reddish reddish reddish reddish Buffer tablets pH 6.8 - for preparing buffer solution acc. to WEISE for the
staining of blood smears
Buffer tablets pH 7.2 - for preparing buffer solution acc. to WEISE for
staining of blood smears
can be used until the stated expiry date.
After first opening of the bottle, the contents can be used up to the stated
expiry date when stored at +5 °C to +30 °C.
The bottles must be kept tightly closed at all times.
The buffer solution can be used for at least 4 weeks when kept in a tightly
closed glass bottle.

Capacity
1 buffer tablet is sufficient for the preparation of 1 l buffer solution acc. to
WEISE.
Additional instructions Literature
For professional use only. 1. H
 istological & Histochemical Methods, J. A. Kiernan,1990, Pergamon
In order to avoid errors, the application must be carried out by qualified Press, Second Edition
personnel only. 2. R
 omeis - Mikroskopische Technik, Editors: Maria Mulisch, Ulrich Welsch,
National guidelines for work safety and quality assurance must be followed. 2015, Springer Spektrum, 19. Auflage
Microscopes equipped according to the standard must be used.
If necessary use a standard centrifuge suitable for medical diagnostic labo- 3. K
 linische Hämatologie, Herbert Begemann, 1975, Georg Thieme Verlag,
ratory. 2. Auflage

Protection against infection

Effective measures must be taken to protect against infection in line with
laboratory guidelines.

Instructions for disposal

The package must be disposed of in accordance with the current disposal
guidelines.
Used solutions and solutions that are past their shelf-life must be disposed
of as special waste in accordance with local guidelines. Information on dis-
posal can be obtained under the Quick Link “Hints for Disposal of
Microscopy Products” at www.microscopy-products.com. Within the EU
the currently applicable REGULATION (EC) No 1272/2008 on classification,
labelling and packaging of substances and mixtures, amending and
repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation
(EC) No 1907/2006 applies.

Auxiliary reagents
Cat. No. 100579 DPX new 500 ml
non-aqueous mounting medium
for microscopy
Cat. No. 101383 Wright’s eosin methylene blue solution 100 ml, 500 ml,
for microscopy 2.5 l
Cat. No. 101424 May-Grünwald’s eosine-methylene blue 100 ml, 500 ml,
solution modified 1 l, 2.5 l
for microscopy
Cat. No. 103699 Immersion oil Type N acc. to ISO 8036 100-ml drop-
for microscopy ping bottle
Cat. No. 104699 Immersion oil 100-ml drop-
for microscopy ping bottle,
100 ml, 500 ml
Cat. No. 105387 Leishman’s eosin methylene blue 500 ml
solution modified
for microscopy
Cat. No. 106009 Methanol for analysis EMSURE® 1 l, 2.5 l, 5 l
ACS, ISO, Reag. Ph Eur
Cat. No. 107961 Entellan® new 100 ml, 500 ml,
rapid mounting medium 1l
for microscopy
Cat. No. 109016 Neo-Mount® 100-ml drop-
anhydrous mounting medium ping bottle,
for microscopy 500 ml
Cat. No. 109204 Giemsa’s azur eosin methylene blue 100 ml, 500 ml,
solution 1 l, 2.5 l
for microscopy

Hazard classification
Cat. No. 109468
Cat. No. 111373
Cat. No. 111374
Please observe the hazard classification printed on the label and the infor-
mation given in the safety data sheet.
The safety data sheet is available on the website and on request.

Main components of the product

Cat. No. 109468
Na2HPO4 x 2 H2O 0.63 g/l
KH2PO4 0.31 g/l

Cat No. 111373

Na2HPO4 x 2 H2O 0.2 g/l
KH2PO4 0.8 g/l

Cat. No. 111374

Na2HPO4 x 2 H2O 0.47 g/l
KH2PO4 0.47 g/l
Consult instructions Manufacturer Catalog number Batch code
Other IVD products for use
Cat. No. 111674 Hemacolor Rapid staining of 1 set
®

blood smear
staining set for microscopy

General remark Caution, consult Use by Temperature

accompanying documents YYYY-MM-DD limitation
If during the use of this device or as a result of its use, a serious incident
has occurred, please report it to the manufacturer and/or its authorised
representative and to your national authority.
Status: 2020-Oct-01
Merck KGaA, 64271 Darmstadt, Germany,
Tel. +49(0)6151 72-2440
www.microscopy-products.com
EMD Millipore Corporation, 400 Summit Drive
Burlington MA 01803, USA, Tel. +1-978-715-4321
Sigma-Aldrich Canada Co. or Millipore (Canada) Ltd.
2149 Winston Park, Dr. Oakville, Ontario, L6H 6J8
Phone: +1 800-565-1400