SU787455A1 - Method of determining living cell biomass - Google Patents

Description

(54) METHOD FOR DETERMINATION OF LIVING BIOMASS

one

The invention relates to the microbiological industry and can be used to determine biomass in liquid media.

There are known methods for determining the biomass of microorganisms, including direct counting of cells under a microscope (in counting chambers, on fixed colored smears, on membrane Filters), extraction and О taking into account seeding on dense (Koch's plate method). ) culture media l.

The disadvantages of these methods are the consideration of both living and dead cells in 15 culture medium, the duration of the methods.

The closest technical solution to the proposed effect achieved is the method of determining the 20 biomass of living cells in a culture medium by introducing a dye into it and then counting the stained cells under a microscope 2,

The disadvantage of the method is that it takes a long time,

The purpose of the invention is to accelerate and simplify the method.

The goal is achieved by IT / T, after adding the dye cell 30

CELL

precipitated from the culture medium, and the counting of living cells is carried out by the amount of dye adsorbed by them. For this test, a certain amount of culture medium (a certain weight for a culture medium containing foam, or a specific volume for a culture medium that does not contain foam) is stained with a thickened amount of in vivo dye, which is known to be larger than necessary for staining living cells, the cells are mixed and precipitated, Without destroying them, using known methods, then the number of LIVE cells is determined by the intensity of the dyeing of the liquid using an in vivo dye.

The color intensity can be determined using a photoelectric colorimeter or other methods.

The color intensity of an in vivo fluid sample of a culture medium uniquely depends only on the number of living cells, since the in vivo dye is consumed only for staining of living cells. The less the intensity of the color of the liquid, the more in the sample of living cells, and the more the intensity of the color

fluid, the less live cells in the sample.

Example. The biomass of living Candida tropical is cells cultured on hydrolyzed media is determined.

PEEUBE of culture medium was taken Aesy 20 g. 5 ml of a 1% aqueous solution of methylene blue was added to it. This mixture is stirred for 5 seconds and 5 ml of Flemming liquid are added with stirring. After 10 s, stirring is stopped. The medium is allowed to settle for 30 seconds - the cells precipitate. According to the intensity of the color of the liquid, the biomass of living cells is determined by using a photometer and a photocolorimeter specially calibrated for this purpose.

The use of the proposed method for determining the biomass of living cells provides an increase in the efficiency of determining the biomass of living cells compared to the existing ones, which makes it possible to use automatic devices, such as photo calorimeters, (This will greatly improve the quality of microorganism culture control.)

Since the proposed method is technologically simple, on its basis it is possible to create an automatic device.

Claims (2)

1. Workshop on Microbiology. Under pej ;. N. p. Yegorova. Tutorial . Moscow, Moscow State University, 1976, p. 57. 2. Methods of experimental mycology. Ed. V.I. Bilay. K., Naukova Dumka, 1973, p. 56.