Cell culture microchambers in a grid matrix sandwiched between a

Chemistry: molecular biology and microbiology – Apparatus – Bioreactor

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Details

435174, 435176, 435180, 435383, 435395, 435401, 4352891, 4353052, 4353053, C12M 300, C12N 500, C12N 1100, C12N 1114

Patent

active

060371714

DESCRIPTION:

BRIEF SUMMARY
FIELD AND BACKGROUND OF THE INVENTION

The present invention relates to an arrangement of habitats, separated from one another, of microscopic dimensions, a method for producing such arrangement as well as methods for investigations and measurements with the aid of this arrangement.
The arrangement described in the present invention of habitats of microscopic dimensions, laid out with microscopic precision on a planar support and separated from one another, is especially suited for the aseptic culture and observation of single cells and their monoclonal descendants as well as also for the culture and study of other small colonies such as bacteria, human, animal or plant cell populations, tissue cultures, mammalian or plant embryos, insects or nematodes at different developmental stages.
To do so, the cells or the other biological samples cited above are placed individually or in relevant combinations into a "pond" disposed on a flat, precisely prepared base; for example a semiconductor, quartz, glass, synthetic or metal base, and covered with a thin semipermeable sheeting, which pond is thin and of extremely small dimensions and which thus forms a sterile microlaboratory. Into each such pond which forms a boxlike closable microchamber, concentrations, adapted to the particular emplaced culture, are added of, for example, buffer components, oxygen, nutrient substances, protective antibiotics, active substances such as pharmacological agents, analytically useful reagents such as for example fluorescent coloring agents etc, by means of penetration of culture media through the semipermeable closable cover sheeting. The supply of, for example, growth factors or other macromolecular medium-specific substances, depending on the selected pore size of the sheeting, can be ensured either before or after the closing of the chamber respectively the pond. It is therein advantageous, for example to place patient sera and expensive biomolecule-preparations into the ponds before closing it in order to minimize its use and to be able to control the concentration in the chambers individually.
The described arrangement provides good conditions for the individual handling and the microscopic observation of the cultures generated therein with respect to their individual properties for example for the controlled exposition in the presence of physical, chemical or other agents and for the optical and physicochemical monitoring and characterization as well as for the final intravital or postmortal storage. This storage can take place for example through deep-freezing or after chemical fixing.
In genetics, microbiology, immunology, oncology, radiobiology, pharmacology, embryology, zoology and botany the need exists for the individual culture of individual cells or multicell samples, in order to observe the growth and the properties of the cultures developing therein, to document them or to preserve them alone or fixed. The invention therefore comprises, in addition to the cited arrangement, respectively its preparation, further the adaptation of the described structures and corresponding measuring parameters in view of embedding the new technology in the current biomedical and biotechnical research and development, as well as for the needs of the intended important applications in clinical diagnostic and therapeutic planning, for the so-called "gene-technology engineering" and applied toxicology or environmental hygiene or pharmacology.
Because the cover sheeting of the ponds can be impermeable to bacteria, viruses and other biological contaminants the microchambers present can be used as sterile laboratories for biological cells. A problem presents itself in the exchange of nutrients and other substances which is only limited via the cover sheeting. With the suggested construction and the tiny cell mass of the individual cultures this exchange problem can, however, be eliminated or reduced.


SUMMARY OF THE INVENTION

It is a task of the present invention to create an arrangement by means of which, to the highest extent independe

REFERENCES:
patent: 4514499 (1985-04-01), Noll
patent: 4693983 (1987-09-01), Davies et al.
patent: 5726060 (1998-03-01), Bridges

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