Copyright Goldenlab International
PRODUCT
TM
Principle

The SPERM STAIN can clearly differentiate the morphological structures of the spermatozoa for a perfect functional assessment. The SPERM STAIN constitutes a system to differentiate the blood cell like components and the spermatozoa as well. The system combines the great polychromic results of classical methods such as May Grunwald or Giemsa but with a really fast method executed in only 15 seconds. 

Product characteristics

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

Staining procedure

1- Fix the smear by immersion in the Working Reagent Nr. 1 ;  5 times, 1 second each. Let drain.
2- Stain the smear by immersion in the Working Reagent Nr. 2 ; 5 times, 1 second each. Let drain again.
3- Stain the smear by immersion in the working Reagent Nr. 3 ; 5 times, 1 second each.

Working reagents

  1. Working reagent Nr. 1 Dilute the vial of SPERM STAIN NR  1 to 1000 ml. with Methanol (we recommend a minimum purity of 99.5% and a water content lower than 0.1%)
  2. Working reagent Nr. 2 Dilute the content of SPERM STAIN NR 2 with 900 ml of deionised  water. Please, use high quality deionized water, no tap water.
  3. Working reagent Nr. 3 Dilute the content of SPERM STAIN NR 3 with 900 ml of deionised  water. Please, use high quality deionized water, no tap water.

Storage and stability

The components of the kit, stored at room temperature (15 - 25C) will remain stable until the expiration date stated on the label. The working reagents are stable at room temperature (15 25 C) for a minimum of 3 months when reagents contamination or excessive evaporation are avoided.

Caution
Reagent n 1, as a methanolic solution is flammable and toxic by contact, inhalation and ingestion.  Handle with care. Waste products must be handled attending your local regulations.
Sample peraparation

Sperm samples: prepare the smear with 15 l of fresh sperm on a standard glass slide. Leave it for air drying at least 10 minutes. It is recommended to make a thin and homogeneous spreading of the sample for the best fixation of the dyes and avoiding blurring hypercolorations.

Rinse the smears gently with deionized water and leave them for air-drying.

In order to prevent deterioration of the smears by the immersion oil coversliping is recommend with an appropriate mounting media such as DPX, Eukitt . Use two drops and a standard glass coverslip ( 50 x 22 mm ).

Remarks

Staining intensity can be modified by varying the number of the immersions in solutions 2 and 3, depending on what color is preferred to be emphasized. Curettes with stain shall always be stored capped, specially 1 in order to avoid undesirable evaporations that could promote color deviations from the usual stainings.
Results

Sperm head:  Homogeneous dark violet Sperm acrosome:  Pale violet, clearer then the head colour Midpiece and tail:  dark violet Background:  Pale pink