[Alginate gel microchip for real-time monitoring of intracellular processes in bacterial and yeast cells].
A method of alginate-based hydrogel cell microchip manufacturing is proposed. The development of mild conditions for cell immobilization in microvolumes of non-toxic alginate gel allows extending the range of microorganisms used. Different approaches to cell analysis using microchip have been approved in pilot studies. By the example of Escherichia coli, Bordetella bronchiseptica and Saccharomyces cerevisiae it is shown that cell microchip can be successfully applied for monitoring of nucleic acid and protein synthesis in growing cells simultaneously using two fluorescent dyes. The influence of chloramphenicol on the nucleic acids and protein synthesis in five bacterial strains has been studied on the microchip. The microchip was also applied for the analysis of inducible fluorescent protein EGFP synthesis in E. coli cells, the correlation between the level of EGFP synthesis and concentration of the inductor in the medium has been established.
Fesenko
D O
DO
Nasedkina
T V
TV
Chudinov
A V
AV
Prokopenko
D V
DV
Iurasov
R A
RA
Zasedatelev
A S
AS
D000464:Alginates; D001426:Bacterial Proteins / Q000096:biosynthesis; D016950:Bordetella bronchiseptica / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D002701:Chloramphenicol / Q000494:pharmacology; D004926:Escherichia coli / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D005656:Fungal Proteins / Q000096:biosynthesis; D005782:Gels; D020723:Glucuronic Acid; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006603:Hexuronic Acids; D009696:Nucleic Acids / Q000096:biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D046888:Tissue Array Analysis / Q000379:methods
D000464:Alginates; D001426:Bacterial Proteins; D005656:Fungal Proteins; D005782:Gels; D006603:Hexuronic Acids; D009696:Nucleic Acids; D011500:Protein Synthesis Inhibitors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D002701:Chloramphenicol; D020723:Glucuronic Acid