Plant Callose Staining Solution (Aniline Blue Fluorescence Method)

Cat. No.: P1511523

GRADE & PURITY BioReagent, Biological Stain, for Fluorescence analysis, for Microscopy

Overview

  Callose is a type of non-cellulosic polysaccharide composed of glucose residues linked in a manner distinct from cellulose. It plays a crucial regulatory role in key biological processes such as plant sieve tube metabolism and gametophyte development. The processes of its synthesis and degradation directly impact normal plant growth and metabolism, making it a significant subject in plant research.

  Plant Callose Staining Solution (Aniline Blue Fluorescence Method) is prepared based on the principle of specific binding between aniline blue and plant callose, which emits a yellow-green fluorescence under specific wavelengths. This method is simple and fast to operate. This product is for research use only and is not suitable for clinical diagnosis or other non-research purposes.

Materials to Be Prepared by User

• AAF Fixative Solution or Carnoy's Fixative Solution, 50% Ethanol, Absolute Ethanol, 1× PBS, Glycerol-Gelatin Mounting Medium or Anti-Fade Mounting Medium
• Fresh plant tissue 

Procedure (For Reference Only)

1. Prepare the Aniline Blue Working Solution: Mix the Aniline Blue Staining Solution and Aniline Blue Diluent in equal proportions. Prepare immediately before use.

2. Cut fresh plant tissue into slices approximately 2 mm thick. For plant leaves, it is recommended to cut them into pieces about 1 cm × 2 cm. Tender plant leaves can be directly immersed in AAF Fixative Solution or Carnoy's Fixative Solution for 24 hours.

3. Rinse the tissue twice with absolute ethanol, 1 minute each time. Then transfer and store the tissue in at least 10 times its volume of 100% ethanol.

4. Before staining, retrieve the tissue and immerse it in 50% ethanol for equilibration for 30 minutes. Remove and allow to drain slightly.

5. Subsequently, immerse the tissue in 1× PBS for equilibration for 30 minutes. Remove and allow to drain slightly.

6. Apply the Aniline Blue Working Solution dropwise onto the tissue section or immerse the section in the working solution. Stain at room temperature, protected from light, for 1~2 hours.

7. Place 25 μL of aqueous Glycerol-Gelatin Mounting Medium or Anti-Fade Mounting Medium on a glass slide. Carefully transfer the stained tissue onto the slide, and add a small additional amount of mounting medium before coverslipping.

8. Observe under a fluorescence microscope using UV excitation light (excitation wavelength 330~385 nm (BP), emission wavelength 425 nm (LP)). Use an unstained section as a control.

Staining Results

  Callose appears as yellow-green fluorescence under UV light, with no background staining.

Notes

1. When conditions permit, it is recommended to subject the plant tissue immersed in fixative to negative pressure/vacuum treatment for 20 minutes to aid fixative penetration.

2. If the experiment is not to be performed immediately or a large number of samples need to be processed simultaneously, samples stored in 100% ethanol can be kept at 2-8°C for at least one week.

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

4. The active components of the staining solution are prone to decomposition. It is recommended to prepare the working solution immediately before use and use it within 3 hours. A slight color fading of the reagent during use is a normal phenomenon.

5. Fluorescent staining is susceptible to photobleaching. Take care to avoid ambient light other than the excitation light during staining, transfer, and observation.

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

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

Storage and Shipping

Storage

Protected from light,Room temperature

Shipped In

Normal

Stability And Storage

Store at room temperature long term (12 months). Store in the dark.

Contents & Storage

P1511523	Component	2×25 mL	Storage
P1511523A	Aniline Blue Staining Solution	25 mL	RT. Store in the dark.
P1511523B	Aniline Blue Diluent	25 mL	RT. Store in the dark.