SPERM STAIN
3 Steps fast staining. For "in vitro" diagnostic
Principle Staining procedure
1- Fix the smear by immersion in the Working Reagent
The SPERM STAIN constitutes a system to differentiate the
Nr. 1 ; 5 times, 1 second each. Let drain.
morphological structures of the spermatozoa for a perfect functional
assessment. It can also be used for the differential staining of blood
2- Stain the smear by immersion in the Working
cells in semen. The stains which make up the SPERM STAIN
Reagent Nr. 2 ; 5 times, 1 second each.
combine polychromy and the quality of classic cytology staining
Let drain again.
methods (May-Grünwald, Giemsa, Wright) with a very quick
execution (time of just 15 seconds). The technique is performed by
3- Stain the smear by immersion in the working
immersion in the staining solutions.
Reagent Nr. 3 ; 5 times, 1 second each.
Product characteristics Rinse the smears gently with deionised water and leave them
The SPERM STAIN system is based on the original Romanowsky
staining method for differential staining of several cellular structures.
Reagents
Kit 3 x 100 ml. (Ref.99 03 85 ). Content:
1. SPERM STAIN NR. 1 1 x 10 ml. (Ref. 99 03 95).
Hexamethyl-p-roseniline methanolic solution. Handle with care.
2. SPERM STAIN NR. 2 1 x 100 ml. (Ref. 99 03 96).
Xanthene buffered solution.
3. SPERM STAIN NR. 3 1 x 100 ml. (Ref. 99 03 97.
Thiazine buffered solution
Working reagents for air-drying.
In order to prevent deterioration of the smears by the
1. Working reagent Nr. 1 immersion oil coversliping is recommend with an appropriate
Dilute the vial of SPERM STAIN NR 1 to 1000 ml. with mounting media such as DPX, Eukitt ®. Use two drops and a
Methanol (we recommend a minimum purity of 99.5% and a standard glass coverslip ( 50 x 22 mm ).
water content lower than 0.1%)
2. Working reagent Nr. 2 Remarks
Dilute the content of SPERM STAIN NR 2 with 900 ml of
deionised water. Please, use high quality deionised water, no Staining intensity can be modified by varying the number of
tap water. the immersions in solutions 2 and 3, depending on what colour
is preferred to be emphasized.
3. Working reagent Nr. 3
Curettes with stain shall always be stored capped, specially 1
Dilute the content of SPERM STAIN NR 3 with 900 ml of
in order to avoid undesirable evaporations that could promote
deionised water. Please, use high quality deionised water, no
colour deviations from the usual stainings.
tap water.
Results
Storage and stability Sperm head: Dark violet
Sperm acrosome: Pale violet, clearer then the head colour
The components of the kit, stored at room temperature (15 - 25º C) Midpiece and tail: dark violet
will remain stable until the expiration date stated on the label. The
Background: Pale pink
working reagents are stable at room temperature (15 – 25 º C) for a
minimum of 3 months when reagents contamination or excessive Red blood cells: Pale or deep pink
evaporation are avoided. Neutrophils: Dark blue nucleus. Pink cytoplasm with red-violet
Caution granulations
Reagent nº 1, as a methanolic solution is flammable and toxic by Eosinophils: Blue nucleus. Blue cytoplasm with red or red-
contact, inhalation and ingestion. Handle with care. Waste orange granules
products must be handled attending your local regulations. Basophils: Dark blue or purple nucleus. Purple, almost black,
granules
Lymphocytes: Violet nucleus. Sky blue cytoplasm
Sample preparation Monocytes: Very pale violet nucleus. Sky blue cytoplasm
Sperm samples: prepare the smear with 15 µl of fresh sperm on a References
standard glass slide. Leave it for air drying at least 10 minutes. It is
recommended to make a thin and homogeneous spreading of the Gurr,E. (1965) "The rational use of dyes in Biology", p. 115.
Leonard Hill, London.
sample for the best fixation of the dyes and avoiding blurring
hypercolorations. Gurr,E. (1971) "Synthetic dyes in Biology, Medicine and
Chemistry". Academic Press. London & New York
Maree, L.; du Plessis, S.S.; Menkvels, R. and van der Horst, G.
Human Reproduction, 25 (6), 1369 – 1382 (2010).
MICROPTIC S.L.
Av. Josep Tarradellas, 8, 1º 6ª
08029 - Barcelona – Spain [Link]
Tel: +34 93 419 29 10
Manufactured by
QUIMICA CLINICA APLICADA S.A. for MICROPTIC S.L.
A 7 Km 1081 – Apdo. 20 - 43870 AMPOSTA / España Rev: 19/12/2017
