Research resource center for molecular and cell technologies (RRC MCT) was open in 2013 at St. Petersburg State University (SPbSU). This center as well as other research resource centers constitute centralized scientific research infrastructure at SPbSU and allow research workers from the university and from other scientific research institutes to access well-developed and expensive technologies in different fields of science.
Research resource center for molecular and cell technologies is the biomedical analytical center aimed at the investigation of the pressing issues in biology and medicine, such as patterns and mechanisms of immune responses, regenerative medicine, biology of aging, cancer, neurodegenerative diseases, etc.
The goal of the RRC MCT is to provide users with the modern biomedical equipment along with the technologies and methods developed on its basis in order to carry out scientific research projects in different fields of biomedicine. Services and technologies provided by the Center are available to internal scientists from SPbSU, researchers from other Russian and foreign universities, research institutes and to commercial enterprises (on the contractual agreements) as well. Specialists of the Center support users with the integral methodological base starting with the help in development of the experimental design of the scientific research projects and finishing with the processing of the results obtained.
The equipment base of the center has more than 50 units of main analytical instruments and sufficient number of routine laboratory equipment for the sample preparation.
Integrated approach to investigations has the priority in modern science especially for the understanding of the fundamental laws of science which can be used in the diagnosis and treatment of diseases, development of the "next generation" immunomodulatory, antimicrobial and other pharmacological agents, search of biologically active substances, development of predictive diagnostics for cognitive abnormalities and neurodegenerative diseases, etc. In order to conveniently carry out such comprehensive investigations, equipment is assembled in 5 different units: the electron microscopy unit, the optical microscopy and cytofluorometry unit, the genomics, metabolomics and proteomics units. Such organization allows to perform complex experiments, especially the ones with strict time limit between stages of sample preparation and analysis when the time is crucial for the success of the whole experiment.
Transmission electron microscopy unit features two microscopes JEOL JEM 1400 and JEM 2100. The unit has all necessary equipment and software for cryo-electron microscopy and electron-microscopic tomography. Pressure freezer Leica HPM‑100 allows researchers to simultaneously stop all the biochemical processes in the sample with complete retention of its morphological structure. Bare-grid plunge freezer Leica EM GP allows to prepare vitrified proteins, viral particles, bacteria for single particle analysis. Cryo‑ultratomy is possible with the help of Leica UC7/FC7 which allows producing ultrathin sections at low temperatures for CEMOVIS and preparation of the material for immuno-gold labeling according to Tokuyasu method for the location identification of the investigated biomolecules on an ultrathin section with the high resolution.
Scanning electron microscopy unit provides scanning electron microscope FE SEM Tescan MIRA3, equipped with 4 axis LN2 cooled stage and 5 axis room temperature stage. Sample preparation equipment includes critical point dryer, metal and carbon coaters, specialized device for freeze-fracture, DARS coating Leica BAF-060 and vacuum cryo transfer shuttle Leica VCT-100.
Advanced light microscopy unit includes two true confocal scanning systems Leica TCS SP 5II, which allow obtaining series of optical “sections” for 3D-reconstruction, as well as performing other specialized techniques. The system features STED‑CW module for super resolution, which provides information on the physiology of a living cell not only at the high spatial resolution, up to 80 nm, but also at the high temporal resolution, up to 18 frames per second. Microscope is also equipped with PicoQuant PicoHarp 300 for analysis of single molecules, FLIM and FC(C)S allowing to explore interaction of biomolecules in living cells under various external influences.
Confocal system Zeiss Cell Observer SD based on the Nipkow disk allows studying fast occurring physiological processes in cells and conducting long-term experiments with living cells due to the low photo-toxicity.
Non-contact laser micro-dissection complex Leica LMD 7000 enables fully automated dissection of tissues with maximum purity being important for the obtaining nucleic acid and protein samples from heterogeneous tissue slices. The complex can also be used to dissect living cells or cell groups according to their fluorescence.
Despite the rapid development of molecular biology approach that helps to answer the questions of "what" and "how" is happening in the experiment, the modern biological technology continues to improve histological methods in order to answer the question "where" the changes are localized. Given the complexity of interaction between cells in tissues, the localization of cellular changes enables to obtain new data necessary for biomedical research. The trend of the last few decades is to obtain accurate quantitative data being achieved by standardization of sample preparation steps. That is why the resource center has automated tissue processing equipment including Tecan GenePaint 150 for automated immunohistochemical staining and FISH.
Sorting cytofluorometer BD FACSAria III, allows rapid acquisition of information about cell internal structure, size, presence of fluorescent labels and sorting the cells of interest accordingly.
Molecular biology unit is designed to solve problems of proteomics, genomics and metabolomics. To cover such a broad range of tasks equipment installed in these units supports a number of sample preparation, purification and concentration techniques, including fractioning by chromatography, one-dimensional and two-dimensional electrophoresis, immunoblotting, preparative electrophoresis.
Proteomics is the study of protein molecules, and their amino acid sequences, spatial structure, post-translational modifications. Proteomics unit provides mass spectrometry analysis with Bruker Ultrafletreme and Agilent 6538 mass spectrometers. The equipment allows determining amino acid sequences of proteins or peptides in a complex mixtures, detection and identification of biomarkers of various organic kinds, conducting molecular imaging of tissues, and identification of microorganisms. Biomolecular interactions could be characterized by BioRad ProteOn 36 system, allowing ranking antibody epitope mapping, studying the interactions of proteins with other proteins, small molecules and nucleic acids. Proteome of two different cell populations or populations grown in different environment could be compared in order to find the up- and down-regulated proteins by DIGE and subsequent scanning resulting 2D gel with laser scanner GE Typhoon 9500FLA.
Genomics is the study of the genetic code in a broad sense. It consists of the structural genomics investigating the nucleotide sequence, the presence of their modifications, interactions with proteins, functional genomics investigating the relationship between the organism's genome and its phenotype, and systematic genomics investigating the use of specific sequences of the DNA in order to clarify the taxonomic position of living organisms. Genomics unit includes sequencing and PCR services. Capillary based Sanger sequencing can be performed on ABI Prism 310 and 3500xl machines. New generation sequencers include Roche GS Junior and ABI Ion Torrent. Single nucleotide polymorphism, number of copy variations and high-performance real-time analysis can be done on Fluidigm BioMark HD, with precise copy number variation analysis being done on BioRad ddPCR.
Metabolomics is currently considered to be one of the most promising areas of molecular techniques in the field of the system biology. Biochemical state of the system, describing metabolite profile, can act as an alternative description of the biological object. Metabolite profiling can provide very useful information on the state and behavior of complex dynamic systems which, in fact, all biological objects are. Metabolomics unit is equipped with the gas chromatography-mass spectrometry system LECO Pegasus 4D and liquid chromatography-mass spectrometry system Agilent 6538 UHD.
The establishment of the RRC "Molecular and cell technologies" was intervened in respect to the interests of scientific research institutes of the Russian Academy of Sciences.
St. Petersburg State Polytechnical University
From July to September 2012 and from December to the end of 2012 Center is working in a test mode, using a small fraction of its equipment. During this period our services were used by
6 Ph.D. students,
25 employees of St.Petersburg State University.
Qualification works were carried out with the help of our equipment
10 (Bachelor, Diploma and Master theses),
Users of the RRC MCT submitted for publication 6 articles
Employees of the Center received 9 certificates.
First contracts with external users are prepared to be signed upon opening of the center.