Calcofluor White (CFW), also known as fluorescent brightener, is a non-specific fluorescent brightening dye. The cell walls of fungi (such as algae) are rich in β-polysaccharides including chitin and cellulose, which can bind to the fluorochrome in the fluorescent brightener to form complexes. Under a fluorescence microscope, specific fluorescence can be observed when excited by a corresponding excitation light source. In plant biology, Calcofluor White is often used to visualize the changes in cell wall formation during the cell wall biosynthesis of algae and higher plants, as well as the identification of protoplast cells.
Fungal Fluorescence Staining Solution (One Step) is mainly composed of fluorescent brightener, Evans blue, potassium hydroxide, dimethyl sulfoxide (DMSO), glycerol, and deionized water. Among these components, the fluorescent brightener can bind to polysaccharides to produce fluorescence; Evans blue can suppress the background fluorescence staining; potassium hydroxide can dissolve cutin, facilitating clearer observation of fungal structures; glycerol has a certain anti-fluorescence quenching effect. As a novel rapid fungal detection stain, this product can be used for fluorescent staining examination of various suspected fungal infections, such as tinea manuum, tinea pedis, tinea versicolor, and pulmonary fungal infections, enabling rapid qualitative detection of fungi under a fluorescence microscope. In addition, this product can also detect Pneumocystis carinii cysts, parasites (e.g., Plasmodium), as well as various bacteria. It can fluorescently label the structural boundaries of these substances, allowing for clear differentiation and detection. This reagent is intended for research use only and is not suitable for clinical diagnosis or any other purposes.
Materials to Be Prepared by User:
1. Fresh or frozen clinical samples; tissue samples embedded in paraffin or glycol methacrylate (GMA)
2. Glass slides, cover slips, fluorescence microscope
Operating Procedures (For Reference Only):
1. Add 1 drop of the staining solution directly onto the sample, ensuring the entire specimen is covered or submerged. Incubate for 2 minutes.
2. Place a cover slip over the sample. Press gently with a cotton swab or absorbent paper to flatten the specimen into a thin layer, then absorb the excess staining solution.
3. Observe the results directly under a fluorescence microscope with an excitation wavelength of 340 nm–380 nm and an emission wavelength of 440 nm.
Staining Results:
Positive: The sample exhibits strong or weak blue-green fluorescence under the fluorescence microscope, and the morphological features are identifiable.
Negative: The morphological features are unidentifiable, or the sample shows extremely weak fluorescence or no fluorescence at all.
Precautions:
1. Personal protective measures should be taken prior to the experiment, as the test samples may carry potential infectivity.
2. Always recap the reagent container immediately after each use to prevent contamination, which could otherwise compromise the staining results.
3. Contaminated waste materials must be disposed of in accordance with national environmental and health safety regulations.
4. Please read the instruction manual thoroughly before use, and make sure to use the reagent within its expiration date.
5. Store the reagent away from extreme high or low temperatures; do not freeze it, and avoid direct sunlight exposure.
6. For your safety and health, wear a lab coat and disposable gloves during the operation.
7. Use the reagent as soon as possible after opening to avoid affecting the outcomes of subsequent experiments.
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