Technical information for the employment of the fluorescence colouring material MYCOVAL in the medicine

by Werner Müller / Helmut Hund GmbH

The MYCOVAL fluorescence colouring material is a reliable, fast and cheap colouring method used for the diagnostics of mycosen (proof of fungi and fungi pores).

It is suitable for:

• native preparations of skin, hair, nails and mucous membrane reductions,
• histological methods, also in combination with other colouring ways (e.g. HE),
• the high-speed diagnostics within the OI range for the proof or exclusion of a mycose,
• the investigation of body fluids and stool,
• air control for fungi elements (e.g. allergy trips),
• the proof of yeast settlements on mucous membranes in the gynaecology

The MYCOVAL colouring is also optimally suitable for the proof of microspores in the stool.

Comparison between more differently microscopic research methods with the fungi investigation method:

What are the special advantages of the MYCOVAL fluorescence colouring material in the medicine?

• The MYCOVAL colouring is the most certain method used to show fungi (e.g. Candida albicans and other Candida species) and/or to Mikrosporidien in the stool.
• The small expenditure and high diagnostic certainty in most cases (in particular within the range of dermatology and gynaecology) permit an immediate introduction to the therapy to be done.
• The expenditure of time for the evaluation of fluorescence preparations to MYCOVAL colouring is minimumal.
• In many cases the composition of the culture can be passed on the colouring material.
• Additionally to the classical methods of staining (e.g. HE) histological preparations can be coloured with MYCOVAL. In the mixing light (transmitted light + top illumination fluorescence) both the fabric-typical structures and possibly preexisting fungi elements can be recognized with certainty.
• Histological high-speed cuts can be immediately coloured and examined.
• MYCOVAL colouring fades under UV light (Fading) and due to this, the stainings are durable for many years.
• MYCOVAL is admitted as the caustic potash solution, which is inserted to the maceration by solid skin particles, hair and nails. Maceration and colouring in a processing step.
• Microscopic investigations can be accounted for after EBM and/or GOO.
• MYCOVAL are harmless, concerning toxicity, in contrary to some other colouring materials.

Product information for Mycoval

Mycoval:
Mykoval is an exclusive product designation for the Helmut Hund GmbH. It is a colouring material solution for the fluorescence microscopy, used to show fungi, including yeasts.
It also concerns the designations "Uvitex 2b" and "Fungiqual A" admitted colouring solutions, however with improved prescription.

Contents:
<1 % ige solution of 4,4`- to { [4-(3-sulfoanilino)-6-bis(2-hydroxyethyl)amino- 1,3,5-triazin-2-yl ] amino } stilben-2,2`-disulfonic acid tetranatrium salt in standard phosphate buffer.
Penetration mediator Dimethylsulfoxid < as 5% (to 15% in permitted for use in some cosmetical goods)

Principle:
Mykoval reacts with chitin and cellulose in the cell wall of the fungi and fluorescences when stimulated by light from the close UV spectrum and the blue range.

Durability:
6 months at room temperature, must be kept away from daylight and UV Radiation.

Note: Mycoval is suitable only for the in-Vitro analysis.

Fungi proof in dermatological practice.

The material which can be examined is taken as usual. Thin skin particles or reductions are moistened on a slide with a drop Mycoval and after approx.. 5 minutes with a cover glass covers. The surplus liquid is squeezed off with flow paper. Afterwards the preparation can be examined immediately - or arbitrarily later.

Stronger striking or nail particle and hair are macerate first with caustic potash solution 10 %-industrial union. This is made best in a damp chamber and can take place e.g. over night. Mycoval can be added to caustic potash solution immediately or before the investigation. If the preparations are to be durably conserved, then one must prevent a draining by bordering the cover glass with nagellack. The Mycoval staining is durable for many years.

Proof of fungi in the vagina area

Yeasts, in particular Candida kinds often settle the mucous membranes in the vagina area. Every now and then such a settlement remains to a large extent symptom-freely, often develops however for the concerning very unpleasant symptoms such as itching, fluorine, etc. and it occurs an unwanted shift of the vaginal flora.

The diagnosis is easy by the proof of yeast fungi to lead.

The reductions are covered on the slide with a drop Mycoval, after approx.. 5 minutes a cover glass is presented, which surplus liquid with flow paper examines immediately squeezed off and the preparation under the fluorescence microscope. Yeast fungi light up clearly cyan and are unmistakable to be differentiated, the background are weakly greenish colored or black. Epithelium cells are still recognizable.

Instruction Sheet for the use of Mycoval

1. Sections of the tissue
   1. The slide should be covered with Mycoval and incubated for 10-30 min
   2. Wash with PBS (phosphate Buffered saltworks)
   3. Microscopic evaluation
      If necessary before the microscopical analysis a contrast dye can be applied and subsequently a wash-out, as well.
2. Drop on a histological coloured material (e.g. HE).
   1. Cover the coloured material with Mykoval, incubate for 10-30 min.
   2. Wash with PBS
   3. Microscopical analysis
3. Smears
   1. Smears should be air-dried and the microscopical slide should be heat fixed.
   2. The slide should be covered with Mykoval and incubated for 5 min.
   3. Wash with PBS
   4. Microscopical analysis.
4. Skin and Nail supplements
   1. Dandruff and nail splinters maceration
   2. Admit Mykoval and incubate for 10 min.
   3. Squeeze off surplus liquid.
   4. Microscopic evaluation.

The microscopic evaluation takes place in a top illumination fluorescence microscope with an excitation wavelength from 390 - 420 nm and a fluorescence filter with 450 nm. Within the blue range the fungi fluoresce yellowish-green. Within the UV range they fluoresce to white-blue. Mykoval shows only a small fading, a long view time is possible.

Test influence
However, cellulose (e.g. cotton stapels) coloured by Mykoval can be easily differentiated from fungi on the basis of their structure. Mykoval was filled up careful considering hygenic regulations. Contamination is to be avoided.