Growth media

The necessary conditions for an accurate analysis of cell morphology by means of cytologic and immunocytochemical studies are a high quality and adequacy of the smear. The quality of the cytologic diagnosis depends on numerous factors: collection of the material, fixation and staining, the method of material examination, the experience of the cytologist and correct interpretation, correct wording of the diagnosis, and use of additional modern research techniques.

Efforts to avoid such mistakes have resulted in the development of a whole range of new fluid-based technologies (Prep Stein, Cytospin, Novo prep, and so on) aimed at enhancing the quality and adequacy of cytologic samples. However all fluid-based technologies require transport media for the biological material which must contain alcohol. Studies by means of the Cytospin system already after the first experiments demonstrated that it was impossible to use the storage medium suggested by Shandon as it led to complete deformation of both the nucleus and the cytoplasm of tumour cells and hindered the routine cytologic analysis of the obtained material, not to mention the evaluation of immune responses.

This “effect of aggression” is connected with the chemical properties of the suggested medium which consists of Denatured ethanol 10% and Polyethylene glycol 5% and was created for stabilisation and fixation of pathologically untransformed cells of the pavement and columnar epithelium in gynaecological smears. In case of morphologically transformed tumour cells with signs of atypism and dystrophy this composition of the medium is unacceptable for both cytologic and immunocytochemical studies.

This was the reason to search for a new storage medium which could correct this defect. Our department has developed and patented a new composition for a storage medium (application No. 2003113164\15 dated 05.05.03, patent for an invention No. 2246110 “Storage and growth medium for a sample of cells intended for subsequent cytologic and/or immunocytochemical analysis”, decision to issue a patent dated 19.05.04).

All cytospin samples used in cytologic and ICC studies are prepared with the help of this growth and storage medium.

The discovered composition of the growth medium has a number of advantages:
Cells retain their morphological, biochemical and biological properties in full without changing the immune specificity, and it is very important for the immunological analysis.
Cells may be stored and transported in this medium for 1 – 2 days.
The facts that the medium is easy to prepare, its components are quite cheap and it is possible to keep it intact at the temperature of 4ºС for a long time make this medium affordable and appropriate for use.
Our growth and storage medium may be recommended for a fluid-based system for preparation of high-quality cytospin samples in case of subsequent cytologic and immunocytochemical studies.

 

After puncturing cells are placed into an Eppendorf tube with the medium. The exposure takes 10 minutes. Then they are centrifuged and applied by a micropipette onto a biochip.

Analysis day No.\

t°Cof the analysis

Cytogram of serous ovarian cancer. (Х200). Immunocytochemistry, expression СК7. (Х200).

1 day/+23°C

Ovarian cancer.
Puncture sample of the vaginal fornix.

1img.

1day-1 1day-2

3 day/+23°C

1img

2day-1 2day-2

5 day/+23°C

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3day-1 3day-2
7 day/+23°C

1img

4day-1 4day-2