NBT-Stained Sample Preservation Solution (For Plant) - BioReagent,for microscopy,Biological Stain

Cat. No.: N1511472

GRADE & PURITY BioReagent, Biological Stain, for Microscopy

Synonyms

Plant Section Preservation Solution (Post-NBT Staining)

Storage

Room temperature

Shipped In

Normal

Overview

  Plant tissues produce various reactive oxygen species (ROS) under stress conditions. These substances are highly reactive and unstable, and are typically quantified and localized by detecting their downstream products. The superoxide anion is a significant type of ROS (oxygen-containing free radical) that can reduce NBT (nitroblue tetrazolium) to an insoluble blue formazan product, thereby achieving in situ localization of superoxide anions in tissues. This method is also known as NBT staining and is suitable for detecting superoxide anions in living plant tissues.

  The NBT Staining Sample Preservation Solution (for Plant) is specifically designed for preserving plant samples after localized superoxide anion staining. It allows stained samples to be stored stably at room temperature for one week and is primarily suitable for whole-mount staining samples of materials such as young root tips and leaves. After staining, areas with superoxide anion accumulation appear blue to dark blue. This product is for research use only and is not intended for clinical diagnosis or other non-research purposes.

Materials to Be Prepared by User

• Fresh plant leaves or roots, tap water, distilled water, 95% ethanol
• Ultrasonic cleaner, magnetic stirrer, electronic balance, filter paper, camera
• NBT Staining Solution (for Plant Use)

Procedure (For Reference Only)

1. Sample Preparation: Collect stressed (e.g., by heavy metals) plant seedlings or root tips. Rinse briefly with tap water and place on filter paper to remove excess moisture.

2. Staining: Immerse the plant seedlings or root tips in the NBT Staining Solution. Stain at room temperature, protected from light, for 2~6 hours, until positive areas develop a dark blue color, while other areas appear light blue, nearly colorless, or retain the plant's natural color. (Adjust staining time based on plant tenderness and degree of color development.)

3. Destaining (Chlorophyll/Background Removal): Carefully remove the seedlings or leaves with forceps. Rinse by immersing and agitating in distilled water 3~5 times. Place on filter paper to remove excess water, then immerse in 95% ethanol and treat at 40°C for 3~16 hours. The purpose is to remove chlorophyll or the light blue background from the seedlings or leaves. Replace with fresh 95% ethanol several times during this process.

4. Observation and Preservation: Remove the seedlings or leaves with forceps. Rinse by immersing and agitating in distilled water 3~5 times. Place on filter paper to remove excess water. Transfer the sample to an adequate amount of NBT Sample Preservation Solution and soak for 30 minutes. Subsequently, the sample can be removed for photography. The sample can be stored in this solution at room temperature for up to one week.

Notes

1. Once prepared, the NBT staining working solution should be stored at 4°C, protected from light, and used within one month. Prolonged storage may affect color development.

2. Since any external factors may stimulate stress-induced production of superoxide anions in plants, plant samples should be freshly collected and stained as soon as possible. It is recommended to include negative and positive control groups.

3. Photograph the stained samples as soon as possible after staining to preserve the results.

4. For the staining and destaining steps, the following alternative procedure is also recommended: Place the tissue in the staining solution, apply vacuum to -0.1 MPa, maintain the negative pressure for 20~30 minutes, then let it stain at room temperature for 60 minutes. Discard the staining solution. Add 95% ethanol and destain in a water bath at 70~80°C, changing the 95% ethanol every 10 minutes. Destaining can be stopped once the green color of the sample has completely faded.

5. For your safety and health, please wear a lab coat and disposable gloves during operation.

6. Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.

7. Recommended NBT Staining Solution Product Numbers: 

• • N1508542 NBT Staining Solution (0.5mg/mL, pH7.8)
    
  • N1508545 NBT Staining Solution (1mg/mL, pH7.8)

Specifications

Synonyms

Plant Section Preservation Solution (Post-NBT Staining)

Specifications & Purity

BioReagent, for microscopy, Biological Stain

Stability And Storage

Store at room temperature long term (12 months).

Storage

Room temperature

Shipped In

Normal

Grade

Biological Stain, BioReagent, for Microscopy