Subjects

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Molecular probes of biological processes - NBCM186

Title:	Molekulární sondy biologických procesů
Form of teaching:	lecture
Guaranteed by:	Institute of Physics of Charles University (32-FUUK)
Faculty:	Faculty of Mathematics and Physics
Actual:	from 2020
Duration in semesters:	1
Semester:	winter
E-Credits:	3
Hours per week, examination:	winter s.:2/0, Ex [HT]
Capacity:	unlimited
Maximum number of enrolled students:	unlimited
Min. number of students:	unlimited
4EU+:	no
Virtual mobility / capacity:	no
State of the course:	taught
Language:	Czech
Teaching methods:	full-time
Repeated enrollment:	2 / 2 / 2 / 2

Guarantor:	Mgr. Josef Lazar, Ph.D.

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Annotation -

Molecular probes of biological processes.

Last update: Procházka Marek, prof. RNDr., Ph.D. (26.05.2021)

Literature - Czech

J. Lakowicz, Principles of fluorescence spectroscopy, Springer 2007

G. Jung, Fluorescent Proteins I and II, Springer 2011

Kianianmomeni, Optogenetics: Methods and Protocols, Springer 2016

Last update: Lazar Josef, Mgr., Ph.D. (26.05.2021)

Syllabus -

Basic principles of detection of molecular probes - optical (fluorescence, luminiscence, vibrational spectroscopy), spin-based, radioactive, electronmicroscopic, mass-spectrometric

Specific demands of molecular probe applications - in vitro, in fixed cells and tissues, in living cells and tissues, in living organisms; labeling of lipids, nucleic acids, and proteins; instruments for detecting individual molecules and molecular assemblies

Principles of labeling - exogenous (affinity, incl. immunodetection, FlAsH/ReAsH, covalent (click-chemistry, photoaffinity labeling), nucleic acid hybridization (DNA barcoding, DIANA), endogenous (genetically encoded - natural and unnatural amino acids)

Labeling for superresolution optical microscopy - exogenous and genetic labeling, relations to mechanisms of optical superresolution

Fluorescent proteins - spectral and other biophysical properties (structure, reactivity, maturation, photoconversion, immunogenicity, FRET, modification tolerance), non-traditional fluorescent proteins

Self-labeling enzymes - dehalogenases (HALO-tag), methyltrasferases and other alkyltransferases (SNAP-tag, CLIP-tag)

Bioluminiscence - lucipherins and lucipherases, BRET, instrumentation

Raman labels - applications and limitations of linear and non-linear Raman spectroscopy based techniques

Optogenetic approaches - modulation of cell membrane voltage, ion concentration, cellular localization, molecular interactions

Detection of neuronal activity - changes in cell membrane voltage, calcium concentration; detection of activity of neurotransmitters, G-protein signaling and other processes

Labeling in electron, cryoelectron and combined electron/optical microscopy

Last update: Procházka Marek, prof. RNDr., Ph.D. (26.05.2021)