A pitfall in using BODIPY dyes to label lipid droplets for fluorescence microscopy
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BODIPY
Lipid droplet
Photoacoustic microscopy is dominantly sensitive to the endogenous optical absorption, while a fluorescence optical microscopy can detect the fluorescence emission to obtain the image of a sample. To some extent, the physical processes of the 2 methods are opposite, one is absorption and another is emission, but both can be used to image cells. In this article, a simultaneous dual-mode imaging system of photoacoustic microscopy and fluorescence optical microscopy is set up to image tobacco cells. Furthermore, gold nanoparticles, which have a large absorption coefficient and enough fluorescence emission with wavelength of 512 nm, are used to label certain drugs and added to the tobacco cells. Then based on the simultaneous dual-mode microscopy imaging system, the photoacoustic microscopy and fluorescence optical microscopy images of gold nanoparticle-labeled tobacco cells are obtained. The final purpose of this experimental research is to detect if the labeled drugs can enter the cells by the positions of the gold nanoparticles. This will help the experts to deliver organic pesticide more accurately and effectively. The experimental results show that by gold nanoparticle labeling technology, the imaging quality of photoacoustic microscopy and fluorescence optical microscopy can be improved, which indicates that the drugs probably enter the tobacco cells successfully.
Fluorescence-lifetime imaging microscopy
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BODIPY
Lipid droplet
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A zinc(II) chelator bis(pyridin-2-ylmethyl)amine moiety has been incorporated into three different highly water-soluble dyes, 2-formyl-BODIPY, 2,6-diformyl BODIPY, and 2,6-diformyl-1,7-distyryl-BODIPY, at 2-position and 2,6-positions, resulting in three highly water-soluble BODIPY-based fluorescent probes A, B and C for zinc(II) ions. Fluorescent probes A and B display sensitive fluorescent responses with significant fluorescence enhancement to zinc(II) ions at pH 7.0 while fluorescent probe C shows two distinct measurable fluorescent signals at 521 nm and 661 nm, and displays ratiometric responses to zinc(II) ions with fluorescence quenching at 661 nm and fluorescence enhancement at 521 nm. These three fluorescent probes exhibit excellent sensitive and selective responses to zinc(II) ions. Intracellular zinc(II) concentration could be monitored in cancer cells with fluorescent probe C.
BODIPY
Moiety
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Selective plane illumination microscopy (SPIM) and other fluorescence microscopy techniques in which a focused sheet of light serves to illuminate the sample have become increasingly popular in developmental studies. Fluorescence light-sheet microscopy bridges the gap in image quality between fluorescence stereomicroscopy and high-resolution imaging of fixed tissue sections. In addition, high depth penetration, low bleaching and high acquisition speeds make light-sheet microscopy ideally suited for extended time-lapse experiments in live embryos. This review compares the benefits and challenges of light-sheet microscopy with established fluorescence microscopy techniques such as confocal microscopy and discusses the different implementations and applications of this easily adaptable technology.
Photoactivated localization microscopy
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Bright-field microscopy
Live cell imaging
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Video microscopy
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Photoactivated localization microscopy
STED microscopy
Fluorescence-lifetime imaging microscopy
Two-photon excitation microscopy
Digital Holographic Microscopy
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The authors present the results of a comparative microscopic study of 502 sputum smears from the patients of the Republican Tuberculosis Dispensary. Sputum smear microscopy with the Ziehl Neelsen (ZN) method and fluorescence microscopy (FM) using a Mikmed 2 (LOMO) microscope and an Olympus SX microscope with a LED Lumin attachment were compared. The latter allows conversion of a light microscope to a fluorescence one. For 2 months sputum was randomly sampled from patients at the moment of diagnosis and from those who were treated at a clinic. A culture study was used as the gold standard, among other things, to calculate the sensitivity and specificity of different microscopy methods. The sensitivity of ZN microscopy, FM on a Mikmed microscope, FM using the Lumin attachment was 28.5, 52.5, and 72.8%, respectively. The Lumin attachment is a inexpensive, portable device that converts practically all models of light microscopes to fluorescence ones. Its life is as long as 25 years, it requires maintenance and both the routine supply line and a Krona storage battery or similar ones, as well as a solar battery may be used as a source of energy.
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Far-red and near infrared (NIR) emissive dyes have advantages in the development of fluorescent probes and labelling for bio-imaging in living systems since fluorescence in the long-wavelength region would generate minimum photo-toxicity to biological components, deep tissue penetration and minimal background from auto-fluorescence by bio-molecules. BODIPY dyes are attractive due to their excellent photo-physical properties and potential for fluorescence-based sensing and bio-imaging applications. Thus, numerous research papers have emerged to develop BODIPY-based dyes with absorption and emission in the long-wavelength spectral region (650–900 nm). This review summarizes the general strategies to obtain far-red and NIR BODIPYs. Moreover, their applications for fluorescent pH probes and imaging or labelling in living systems are highlighted.
BODIPY
Far-red
Fluorescence-lifetime imaging microscopy
Biological Imaging
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Preface.- Acknowledgements.- Dedication.- Ray Optics, Wave Optics and Imaging System Designing.- Basics of Electromagnetic Theory for Fluorescence Microscopy.- Electric Field Effects in Optical Microscopy Systems.- Quantum Description of Radiation Field and Optical Microscopy.- Molecular Physics of Fluorescence Markers.- Basics of Fluorescence and Photophysics.- General Fluorescence Imaging Techniques.- Multiphoton Fluorescence Microscopy.- Super Resolution Fluorescence Microscopy.- Image Reconstruction Methodologies for Fluorescence Microscopy.- Future Prospective of Fluorescence Microscopy.- Appendix I.- Appendix II.- Appendix III.
Photoactivated localization microscopy
Fluorescence-lifetime imaging microscopy
Fluorescence cross-correlation spectroscopy
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