Pro-Stains™ Malachite Green Solution

Pro-Stains™ Malachite Green Solution is a high-quality staining reagent used in the Acid-Fast staining method for the microscopic detection of mycobacteria in clinical specimens and laboratory cultures. The solution is available in multiple volumes to suit different laboratory needs:

• PL.7030: 500 mL
• PL.7031: 1 L
• PL.7032: 2 L

This solution is specifically formulated for consistent and reliable performance in Ziehl-Neelsen and Kinyoun staining methods, ensuring accurate visualization of acid-fast bacteria, including species of Mycobacterium such as M. tuberculosis.

Key Features

1. High-Quality Formulation:
   • Contains malachite green, a counterstain that provides excellent contrast for non-acid-fast organisms against the red-stained acid-fast bacteria.
2. Versatile Applications:
   • Compatible with both Ziehl-Neelsen (heat-fixed) and Kinyoun (cold) staining techniques.
   • Suitable for routine diagnostics and research applications.
3. Multiple Packaging Sizes:
   • Available in 500 mL, 1 L, and 2 L bottles to accommodate laboratories with varying testing volumes.
4. Reliable Staining Performance:
   • Produces sharp differentiation between acid-fast and non-acid-fast organisms, ensuring high diagnostic accuracy.
5. Meets Diagnostic Standards:
   • Designed for in vitro diagnostic use, meeting the needs of clinical microbiology labs and research institutions.
6. Easy to Use:
   • Ready-to-use formulation eliminates the need for additional preparation.

Composition and Principle

• Malachite Green: Serves as a counterstain in the acid-fast staining process, coloring non-acid-fast bacteria and background materials green, while acid-fast bacteria retain the primary stain (red) due to the high lipid content of their cell walls.

Principle:

In acid-fast staining, the primary stain (carbol fuchsin) penetrates the waxy cell wall of acid-fast bacteria due to heating or the presence of a detergent. Non-acid-fast organisms cannot retain the primary stain after decolorization with acid-alcohol. The counterstain, malachite green, is then applied to highlight the non-acid-fast components, enhancing visualization under a microscope.

Applications and Use Cases

1. Clinical Diagnostics

• Purpose:
  • To identify Mycobacterium tuberculosis, the causative agent of tuberculosis, and other acid-fast bacteria in patient specimens.
• Use Case:
  • A hospital microbiology lab uses Pro-Stains™ Malachite Green Solution as part of Ziehl-Neelsen staining to detect M. tuberculosis in sputum smears.

2. Public Health Surveillance

• Purpose:
  • To monitor and track tuberculosis and other acid-fast bacterial infections in community health programs.
• Use Case:
  • A public health lab screens sputum samples from suspected TB patients using Kinyoun staining with malachite green as the counterstain.

3. Research Applications

• Purpose:
  • To study acid-fast bacteria, including non-tuberculous mycobacteria (NTM), in academic and pharmaceutical research.
• Use Case:
  • A research lab uses Pro-Stains™ Malachite Green Solution to differentiate M. avium from non-acid-fast bacteria in mixed cultures.

4. Environmental Testing

• Purpose:
  • To detect Mycobacterium species in environmental samples such as water and soil.
• Use Case:
  • An environmental lab uses malachite green in Kinyoun staining to identify M. leprae in soil samples from endemic regions.

Step-by-Step Instructions

Ziehl-Neelsen Method (Heat-Based):

1. Prepare a heat-fixed smear on a clean glass slide.
2. Flood the slide with carbol fuchsin (primary stain) and heat gently until steaming. Avoid overheating.
3. Allow to stain for 5–10 minutes.
4. Rinse the slide gently with water.
5. Decolorize with acid-alcohol until no more red color washes out.
6. Rinse with water.
7. Counterstain with Pro-Stains™ Malachite Green Solution for 1–2 minutes.
8. Rinse with water and air-dry.
9. Examine under a microscope using oil immersion (100x objective).

Kinyoun Method (Cold Stain):

1. Prepare a heat-fixed smear on a clean glass slide.
2. Flood the slide with carbol fuchsin and allow to stain for 5–10 minutes without heating.
3. Rinse the slide gently with water.
4. Decolorize with acid-alcohol until no more red color washes out.
5. Rinse with water.
6. Counterstain with Pro-Stains™ Malachite Green Solution for 1–2 minutes.
7. Rinse with water and air-dry.
8. Examine under a microscope using oil immersion (100x objective).

Interpretation of Results

• Acid-Fast Bacteria: Retain the red color of the primary stain (carbol fuchsin) due to their waxy, lipid-rich cell walls.
• Non-Acid-Fast Bacteria: Appear green due to the malachite green counterstain.
• Background Material: Stains green, providing contrast for improved visualization.

Advantages

1. Enhanced Contrast:
   • Clear differentiation between acid-fast and non-acid-fast organisms improves diagnostic accuracy.
2. Reliable Results:
   • High-quality formulation ensures consistent and reproducible staining results.
3. Ease of Use:
   • Ready-to-use solution simplifies workflows in busy laboratories.
4. Flexible Volumes:
   • Available in various sizes to meet the needs of labs with differing workloads.
5. Cost-Effective:
   • Affordable solution for routine acid-fast staining procedures.

Specifications

Precautions

• For in vitro diagnostic use only.
• Use appropriate personal protective equipment (PPE) when handling the reagent.
• Avoid direct contact with skin and eyes. In case of contact, rinse thoroughly with water.
• Dispose of used reagents and materials in accordance with local regulations for hazardous waste.

Storage and Stability

• Store at 15–25°C in a dry environment.
• Keep the container tightly closed to prevent contamination.
• Stable until the expiration date indicated on the label.

Why Choose Pro-Stains™ Malachite Green Solution?

1. Quality Assurance:
   • Reliable and consistent staining results for acid-fast bacteria.
2. Ease of Use:
   • Ready-to-use formulation simplifies laboratory workflows.
3. Versatility:
   • Compatible with Ziehl-Neelsen and Kinyoun staining techniques.
4. Multiple Sizes:
   • Flexible volume options for laboratories of all sizes.
5. Cost-Effective:
   • Affordable solution for routine acid-fast staining needs.

References

1. American Society for Microbiology (ASM): Clinical Microbiology Procedures Handbook.
2. WHO Guidelines: Laboratory diagnosis of tuberculosis by sputum microscopy.
3. CDC Protocols: Acid-Fast Staining Procedures for Mycobacterium detection.

Pro-Stains™ Malachite Green Solution is an essential tool for accurate and reliable acid-fast staining, making it a valuable asset for clinical diagnostics, research, and public health laboratories.