SUBJECT

Experimental Methods in neurobiology I.

lecture

master

Faculty of Science

Biology MSc

2

Semesters 1-4

Autumn/Spring semester

1. History of methodology in neuroscience.
2. Molecular biology techniques: DNA, RNA, protein technologies Molecule detection methods: PCR Microarray, protein, DNA, mRNA, cDNA Gene switching In situ PCR In situ hibridization Histochemistry, basics in immune-histochemistry Gene insertation, gene manipulation in neurons transfection, viral and non-viral gene transfere, genegun, electroporation, gene scilencing (siRNA)
3. Animal models in neurosciences: Application of genetically modified strains. Spontaneous mutation, inbread strains. Controlled gene insertation: transgenic animals, knock out, knock in, conditional knock out, tissue specific and inducible gene modification. Functional enzyme mutants.
4. Optical methods: Normal light and fluorescen microscopy; selection of detectable molecular teg; natural and synthetic fluorescent molecules. Optical slicing: confocals laser scanning microscopy, multi-photon microscopy, spinning microscopy. Computerized image analysis, "sliceing": deconvolution methods, ApoTome Molecule detection: Single molecule tracing microscopy. Protein protein interactions: FRET (fluorescence resonance energy transfer), TIRF (total internal reflection fluorescence), Biachor/OWLS, BRET (Bioluminescence resonance energy transfer) Kinetic studies: time lapse/live cell microscopy Imaging (intracellular Ca2+, pH, membrane potencial changes) Video-microscopy: cell shape- and motility changes FRAP (fluorescence recovery after photobleaching), FLIP (fluorescence lifetime imaging), FCS (fluorescence correlation spectroscopy); SPT (Single particle tracking); intracellular transport of vesicles, photo activated molecules

• G.B. Smejkal, A. Lazarev Separation Methods in Proteomics, Taylor&Francis, NY, 2006

• S. Srivastava Informatics in Proteomics, Taylor&Francis, NY, 2005

• J.H. Gross Mass Spectrometry, Springer, 2004