PROCEDURE FOR EVALUATION OF FRACTION OF HYBRID CELLS AND CELL PRODUCTS BY MEANS OF CONFOCAL MICROSCOPY AND AUTOCLAVABLE ELECTROFUSION CHAMBER

摘要

The subject of the invention belonging to the filed of biomedicine technology is a procedure of evaluation of the fraction of hybrid cells and cell products - hybridoms - in a cell vaccine, which comprises measurement of surface area of colocalized pixels on a picture taken by a multichannel confocal microscope, as well as an autoclavable electrofusion chamber intended for multiple use in preparation of hybridom cells. The procedure for quantification of fraction of hybrid cells enables the evaluation of a hybrid cell fraction by measuring surface areas of colocalised pixels and simultaneous visualisation and documentation of the procedure. The applicability of the procedure is wide also for any cases where fractions of one or several cell types or structures on a picture are determined. The subject of invention is also a respective autoclavable electrofusion chamber, more precisely, a chamber for the fusion of biological cells in the electrical field. The invention refers to the structure of the electrofusion chamber which is intended for multiple application. The chamber is composed of an upper part (1), lower part (2), glass plate (4) with electrodes, o-ring, cover and nut, which all can be autoclaved separately and put together as a sterile electrofusion chamber very easily in a sterile particle-free chamber. Besides, it is possible to change the size of the electrofusion surface by changing individual parts of the electrofusion chamber and thus fuse various volumes of cell suspensions in the chamber. The electrofusion chamber is made of biocompatible materials and is suitable for preparation of hybrid cells for human vaccines.