Gram Fluorescent Staining Probe‌ (AIE) - BioReagent, 10mM

Cat. No.: A1456427

GRADE & PURITY BioReagent, 10mM

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Application

Microbial staining

Overview

  The Gram Fluorescent Staining Probe‌ (AIE) is a small-molecule probe developed based on the Aggregation-Induced Emission (AIE) principle, featuring typical AIE characteristics. It can bind to different structures of various bacterial species and enable rapid differentiation between Gram-negative and Gram-positive bacteria through fluorescence.

Product Characteristics

  The outer membrane of Gram-negative bacteria serves as a barrier, which can effectively prevent AIE fluorescent dyes from entering the bacterial cells. However, the outer membrane can adsorb AIE fluorescent dyes onto the bacterial surface through electrostatic adsorption, causing the bacteria to exhibit green fluorescence under a fluorescence microscope.Gram-positive bacteria lack an outer membrane structure, allowing AIE fluorescent dyes to penetrate the cell membrane and enter the bacterial cells. As a result, Gram-positive bacteria show orange-yellow fluorescence under a fluorescence microscope.

Figure 1. Staining effect of the Gram Fluorescent Staining Probe‌ (AIE) on Staphylococcus aureus and Klebsiella spp. (Green: Klebsiella spp.; Yellow: Staphylococcus aureus)

Product Advantages

1. It enables naked-eye differentiation between Gram-negative and Gram-positive bacteria based on the difference in emitted fluorescence color.

Test Method

1. Preparation of Dye Stock Solution: After brief centrifugation, aliquot the dye stock solution appropriately and store it in the dark at -20°C or lower.
2. Preparation of Dye Staining Solution: Add 1 μL of the dye stock solution to 500 μL of ultrapure water to prepare a staining solution with a concentration of 20 μM.
3. Bacterial Preparation: Take the bacteria cultured overnight, centrifuge them, wash the bacterial pellet 2-3 times with PBS, and concentrate the bacterial solution. It is recommended that the final bacterial concentration have an OD₆₀₀ value of 0.2-1.5.
4. Bacterial Smearing: Drop 2-3 μL of the bacterial solution onto a glass slide and spread the sample evenly with a cotton swab.
5. Staining and Imaging: Drop 20-40 μL of the staining solution onto the glass slide, cover it with a coverslip, and observe the smear using the long-pass B band of a fluorescence microscope (Excitation: 460-490 nm; Emission: >510 nm). Gram-negative bacteria will be stained green, while Gram-positive bacteria will be stained orange.

Precautions

1. Centrifuge briefly before use.
2. Store at -20°C and avoid repeated freezing and thawing.
3. For your safety and health, wear a lab coat and disposable gloves during operation.
4. This product is for research use only.

Specifications

Specifications & Purity

BioReagent, 10mM

Stability And Storage

Store at -20℃ (1 years). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle. Store in the dark.

Storage

Protected from light, Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent