ISSN 2312-3672
Orekhov F., Jablokov A., Skrynnik A. Ybridization of laser-induced spectrofluorescence analysis (lifs), matrix-assisted laser desorption / ionization mass spectrometry (maldi), fluorescence recovery after photobleaching (frap) anf fluorescence loss in photobleaching (flip) microtechnics // Journal of Biomedical Technologies. 2016. № 2. P. 42‒52. DOI: 10.15393/j6.art.2016.3702
Issue № 2 (2016)
Brief Communications
Ybridization of laser-induced spectrofluorescence analysis (lifs), matrix-assisted laser desorption / ionization mass spectrometry (maldi), fluorescence recovery after photobleaching (frap) anf fluorescence loss in photobleaching (flip) microtechnics
| Orekhov Fedor K. | Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia, a.g.jablokow@gmail.com |
| Jablokov Arthur G. | Pirogov Russian National Research Medical University, Moscow, Russia, a.g.jablokow@gmail.com |
| Skrynnik Andrew A. | Institute of Energy Problems of Chemical Physics, Russian Academy of Sciences, Moscow, Russia, a.g.jablokow@gmail.com |
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Matrix-Assisted Laser Desorption / Ionization Mass Spectrometry (MALDI) Laser-Induced Spectrofluorescence Analysis (LIFS) Fluorescence Loss in Photobleaching (FLIP) Fluorescence Recovery After Photobleaching (FRAP) Confocal Laser Scanning Microscopy (CLSM) |
Novel MALDI MS + FLIP approaches for verifying continuity of membranous structures and measurements of nucleus-cytoplasm exchange rates are proposed. Novel approaches for the measurements of lateral diffusion / molecular mobility and bindings using MALDI + FRAP hybridization are proposed. FRAP (Fluorescence Recovery After Photobleaching) is a method for the diffusion kinetics measurements in living cells using fluorescence microscopy which allows to estimate quantitatively the two dimensional lateral diffusion in molecularly thin film containing fluorescent-labeled probes, or for single cell examination (i.e. the study of lateral mobility of cellular molecules). Fluorescence Loss in Photobleaching (FLIP) is a microscopic technique predominantly performed using laser scanning microscopy (e.g. for tagged protein local photobleaching by short, intensive laser excitation on CLSM platform) used for the studies on molecular mobility inside the cells and membranes. MALDI (Matrix-Assisted Laser Desorption / Ionization) is a soft ionization technique used in mass spectrometry, allowing for the analysis of biomolecules (biopolymers such as DNA, proteins, peptides and sugars) and large organic molecules (such as polymers, dendrimers and other macromolecules), which tend to be fragile and fragmented when ionized by more conventional ionization methods (according to encyclopedic definition). The above laser-based technique is readily compatible with MALDI using the same laser beam for MALDI and FLIP. |
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