Christmas Tree stain

Two chemicals are used together to stain sperm cells; Kernechtrot solution will stain the head of the sperm cell a two-tone reddish pink color and the picroindigocarmine solution will stain the tail of the sperm cell a bluish-green color.

Christmas Tree Stain for Sperm

Kernechtrot-Picroindigocarmine Stain (Christmas Tree Stain

• Kernechtrot-Picroindigocarmine (KPIC) differential biological stain is used to assist in the microscopic identification of spermatozoa. The solutions for this procedure can either be purchased or prepared in-house.
• Safety Considerations
• Aluminum sulfate - Caution! Harmful if inhaled, in contact with skin, and if swallowed! Emits toxic fumes under fire conditions!
• Nuclear fast red - Caution! Irritant! Emits toxic fumes under fire conditions!
  
• Saturated picric acid solution - Caution! Toxic! Explosive when dry! Emits toxic fumes under fire conditions!
• Indigocarmine dye - Caution! Harmful if swallowed! Emits toxic fumes under fire conditions!

Equipment

• Flame or heat block

Materials

• Fixative (optional)

Reagents

• Kernechtrot staining solution (KS)
• Picroindigocarmine staining solution (PICS)
• Distilled water
• 95% ethanol or methanol
• Stock Solutions (In-house Preparation)

Equipment

• Filtration apparatus
• 500 ml glass beakers
• Balance
• Spatula
• Glass rod
• Plastic bottles

• Materials
• Filter paper
• Weigh boats or weigh paper

   * Reagents

• Aluminum sulfate
• Nuclear Fast Red
• Distilled water
• Picroindigocarmine dye
• Saturated picric acid solution
* Kernechtrot Solution (KS)
• In a beaker dissolve 5 g of aluminum sulfate in 100 ml of hot
distilled water.
• Immediately add 0.1 g of Nuclear Fast Red and
stir with a glass rod.
• Allow to cool and filter through filter paper.

• Storage

• • The Kernechtrot Solution is stable at room temperature for up to 6 months, but may need to be refiltered after standing.
* Labeling
* Label the bottle as KS with the expiration date and a lot number (the date of preparation followed by the initials of person preparing the solution). * Picroindigocarmine Solution (PICS)
• Dissolve 1 g of Indigocarmine dye in 300 ml of a commercially
purchased saturated solution of picric acid.
• Filter and store.

Storage

• The Picroindigocarmine Solution is stable at room temperature for up to 6 months, but may need to be refiltered after standing.

Labeling

• • Label the bottle as PICS with an expiration date and a lot number (the date of preparation followed by the initials of person preparing the solution).

Christmas Tree Staining Procedure

• Prepare a thin smear of an extract of a suspected semen stain and allow to dry. 
• Fix the smear with a quick flame or fixative, or by placing it on a heat block overnight.
• Add a sufficient amount (2-5 drops) of red reagent to cover the stained portion of the microscope slide.
• Let the slide stand at room temperature for at least 15 minutes.
• Wash off of the slide with a gentle stream of distilled water and drain the slide.
• Add a sufficient amount (2-5 drops) of green reagent to cover the stained portion of the slide.
• Allow green reagent to stain the smear for 5-15 seconds.
• Wash green reagent off of the slide with 95% ethanol or methanol.
• Dry the slide at room temperature.




Jump to: Main Page • Micropedia • Macropedia • Icons • Time Line • History • Life Lessons • Links • Help
Chat rooms • What links here • Copyright info • Contact information • Category:Root