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Cat.No: FS-1651 Datasheet Instruction for Use
Specification Quantities
Price $494.13–823.55
| Product Name | Sperm Morphology Stain Kit (Papanicolaou Method) |
| Catalog No. | FS-1651 |
| Description | The Papanicolaou (Pap) stain is widely used for routine cytological staining. Initially, the Papanicolaou stain was used solely to detect estrogen levels in vaginal epithelium and to identify pathogens such as Candida and Trichomonas in the reproductive tract. When Orange G6 is used in combination with EA36 or EA50, it stains the cytoplasm in vivid shades of green, blue, and pink. Sperm morphology staining kit (Papanicolaou method): Since proteins with different isoelectric points within sperm and cells carry different charges at the same pH, they selectively bind to corresponding dyes and become stained. The nucleus is composed of acidic substances and has a strong affinity for basic dyes; whereas the cytoplasm, conversely, contains basic substances and has a higher affinity for acidic dyes. For cytoplasmic staining, a modified EA50 staining solution specifically designed for sperm staining is used, while nuclear staining employs a proprietary, non-toxic modified hematoxylin solution. This kit is particularly suitable for sperm staining and can also be used for staining cellular samples such as pleural fluid, ascites, and sputum. |
| Test Procedure | Thaw fresh semen specimens that have fully liquefied or those stored at -20°C (to room temperature). If sperm washing is required prior to preparation, follow these steps: (1) Add 0.5 mL of semen and 10 mL of saline to a test tube and mix thoroughly. Centrifuge at 2000 rpm for 10 minutes, then remove most of the supernatant. Gently tap the test tube to allow the sperm clump to resuspend in the remaining saline. If the sperm concentration is too high, adjust the sperm density appropriately with saline. Place 10–20 μL of the sperm suspension onto a clean microscope slide and spread it evenly into a circular sample area. Place the prepared slide horizontally and allow it to air dry naturally. Fix the dried smear in a 95% ethanol solution for 5–10 minutes. Dehydrate using a series of ethanol solutions (80%, 70%, 50%) in descending order, allowing 1 minute at each concentration. Soak or rinse in distilled water or tap water for 1 minute. Stain with hematoxylin solution for 3–5 minutes. Rinse with distilled water for 30 seconds. Differentiate with acidic ethanol for approximately 4–5 seconds; it is recommended to observe under a microscope to ensure adequate differentiation. Blue with the blueing solution for 4 minutes. Rinse with tap water for 2 minutes. Dehydrate with a series of ethanol (50%, 70%, 80%, 95%), 1 minute per step. Stain with Orange G6 solution for 2 minutes. Rinse with 95% ethanol (I) and (II) for 2 minutes each. Stain with modified EA50 solution for 5 minutes. Dehydrate with 95% ethanol (I) and (II) for 1 minute each. Dehydrate with anhydrous ethanol (I) and (II) for 1 minute each. Clear with xylene and mount with neutral resin. |
| Results | Sperm acrosome region—light blue Non-acrosomal region of the head (post-acrosomal region)—dark blue Midpiece—slightly reddish Sperm tail—blue or light red Excess residual cytoplasm, typically located beneath the head or surrounding the midpiece, stains red. |
| Storage and Transportation | Store at room temperature, protected from light. Shelf life: 1 year. |
| Precautions | For your safety and health, please wear a lab coat and disposable gloves when performing this procedure. |
For research use only, not for clinical use.
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