Application Of Laser Scanning Microscopy Research Articles

Suppression of Subpixel Jitter in Resonant Scanning Systems With Phase-locked Sampling.

Resonant scanning is critical to high speed and in vivo imaging in many applications of laser scanning microscopy. However, resonant scanning suffers from well-known image artifacts due to scanner jitter, limiting adoption of high-speed imaging technologies. Here, we introduce a real-time, inexpensive and all electrical method to suppress jitter more than an order of magnitude below the diffraction limit that can be applied to most existing microscope systems with no software changes. By phase-locking imaging to the resonant scanner period, we demonstrate an 86% reduction in pixel jitter, a 15% improvement in point spread function with resonant scanning and show that this approach enables two widely used models of resonant scanners to achieve comparable accuracy to galvanometer scanners running two orders of magnitude slower. Finally, we demonstrate the versatility of this method by retrofitting a commercial two photon microscope and show that this approach enables significant quantitative and qualitative improvements in biological imaging.

Application of laser scanning microscopy for the analysis of oral biofilm dissolution by different endodontic irrigants.

Multi-specie biofilms are highly resistant to antimicrobials due to cellular interactions found in them. The purpose of this study was to evaluate, by confocal laser scanning microscopy, the biofilm dissolution effectiveness of different irrigant solutions on biofilms developed on infected dentin in situ. A total of 120 bovine dentin specimens infected intraorally (30/group) were treated by the following solutions: 2% of chlorhexidine digluconate, 1%, 2.5% and 5.25% of sodium hypochlorite (NaOCl). The solutions were utilized for 5, 15 and 30 min with 2 experimental volumes 500 μL and 1 mL. All the samples were stained using an acridine orange and the biofilm thickness before (control group) and after the experiments were evaluated, utilizing a confocal microscope at ×40. The Mann-Whitney U and the nom-parametric Kruskal-Wallis Dunns tests were utilized to determine the influence of the volume and to perform the comparisons among the groups respectively. The significance level was set at P < 0.05. Statistical differences were not found among the control and the 2% chlorhexidine digluconate groups at any experimental period (P > 0.05). The biofilm dissolution treated with 1% NaOCl was directly proportional to the exposure time (P < 0.05). The higher values of biofilm dissolution were found in 2.5% and 5.25% NaOCl groups (P > 0.05). The higher exposure times and concentrations of NaOCl were not sufficient to dissolve 100% of the biofilm. However, all NaOCl solutions were more effective than 2% chlorhexidine digluconate to dissolve organic matter.

3D imaging of biofilms on implants by detection of scattered light with a scanning laser optical tomograph

Biofilms – communities of microorganisms attached to surfaces – are a constant threat for long-term success in modern implantology. The application of laser scanning microscopy (LSM) has increased the knowledge about microscopic properties of biofilms, whereas a 3D imaging technique for the large scale visualization of bacterial growth and migration on curved and non-transparent surfaces is not realized so far.Towards this goal, we built a scanning laser optical tomography (SLOT) setup detecting scattered laser light to image biofilm on dental implant surfaces. SLOT enables the visualization of living biofilms in 3D by detecting the wavelength-dependent absorption of non-fluorescent stains like e.g. reduced triphenyltetrazolium chloride (TTC) accumulated within metabolically active bacterial cells. Thus, the presented system allows the large scale investigation of vital biofilm structure and in vitro development on cylindrical and non-transparent objects without the need for fluorescent vital staining. We suggest SLOT to be a valuable tool for the structural and volumetric investigation of biofilm formation on implants with sizes up to several millimeters.

Laser scanning microscopy of HTS films and devices (Review Article)

The capabilities of laser scanning microscopy (LSM) as a spatially-resolved method of testing high-Tc superconductivity (HTS) materials and devices are described. The earlier results obtained by the authors are briefly reviewed. Some novel applications of LSM are illustrated, including imaging the HTS responses in rf mode, probing the superconducting properties of HTS single crystals, and development of two-beam laser scanning microscopy. The existence of the phase slip lines mechanism of resistivity in HTS materials is proven by LSM imaging.

Molecular Dynamics in Living Cells Observed by Fluorescence Correlation Spectroscopy with One- and Two-Photon Excitation

Multiphoton excitation (MPE) of fluorescent probes has become an attractive alternative in biological applications of laser scanning microscopy because many problems encountered in spectroscopic measurements of living tissue such as light scattering, autofluorescence, and photodamage can be reduced. The present study investigates the characteristics of two-photon excitation (2PE) in comparison with confocal one-photon excitation (1PE) for intracellular applications of fluorescence correlation spectroscopy (FCS). FCS is an attractive method of measuring molecular concentrations, mobility parameters, chemical kinetics, and fluorescence photophysics. Several FCS applications in mammalian and plant cells are outlined, to illustrate the capabilities of both 1PE and 2PE. Photophysical properties of fluorophores required for quantitative FCS in tissues are analyzed. Measurements in live cells and on cell membranes are feasible with reasonable signal-to-noise ratios, even with fluorophore concentrations as low as the single-molecule level in the sampling volume. Molecular mobilities can be measured over a wide range of characteristic time constants from ∼10 −3 to 10 3 ms. While both excitation alternatives work well for intracellular FCS in thin preparations, 2PE can substantially improve signal quality in turbid preparations like plant cells and deep cell layers in tissue. At comparable signal levels, 2PE minimizes photobleaching in spatially restrictive cellular compartments, thereby preserving long-term signal acquisition.

Application of Laser Scanning Microscopy to Morphological Observations on Drinking in Freshwater Medaka Larvae and Those Exposed to 80% Seawater

Application of Laser Scanning Microscopy to Morphological Observations on Drinking in Freshwater Medaka Larvae and Those Exposed to 80% Seawater

Applications of laser scanning microscopy for analysis of aquatic microhabitats.

Laser scanning microscopy was used 1) to analyze Mn oxide-encrusted biofilms and particles in marine Mn-oxidizing enrichment cultures, 2) to optimize fluorescence in situ hybridization protocols of 16S ribosomal RNA-targeted oligonucleotide signature probes for single bacterial cell identification in particles from a wetland, and 3) to develop a combined immunofluorescence-microautoradiography procedure for analysis of the distribution of 14C-labeled organic compounds and 14C-mineralizing bacteria in groundwater seep sediments. The results demonstrated the wide applicability and benefits of using laser scanning microscopy for analysis of complex microbial assemblages.

Application of Laser Scanning Microscopy to Nucleo‐Cytoplasmic Transport in Single Living Cells

Berichte der Bunsengesellschaft für physikalische ChemieVolume 92, Issue 9 p. 991-993 Article Application of Laser Scanning Microscopy to Nucleo-Cytoplasmic Transport in Single Living Cells Reiner Peters, Reiner Peters Max-Planck-Institut für Biophysik, Kennedyallee 70, 6000 Frankfurt 70, Federal Republic of GermanySearch for more papers by this authorManfred Scholz, Manfred Scholz Max-Planck-Institut für Biophysik, Kennedyallee 70, 6000 Frankfurt 70, Federal Republic of GermanySearch for more papers by this authorClaudia Grosse-Johannböcke, Claudia Grosse-Johannböcke Max-Planck-Institut für Biophysik, Kennedyallee 70, 6000 Frankfurt 70, Federal Republic of GermanySearch for more papers by this author Reiner Peters, Reiner Peters Max-Planck-Institut für Biophysik, Kennedyallee 70, 6000 Frankfurt 70, Federal Republic of GermanySearch for more papers by this authorManfred Scholz, Manfred Scholz Max-Planck-Institut für Biophysik, Kennedyallee 70, 6000 Frankfurt 70, Federal Republic of GermanySearch for more papers by this authorClaudia Grosse-Johannböcke, Claudia Grosse-Johannböcke Max-Planck-Institut für Biophysik, Kennedyallee 70, 6000 Frankfurt 70, Federal Republic of GermanySearch for more papers by this author First published: September 1988 https://doi.org/10.1002/bbpc.198800247AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Volume92, Issue9September 1988Pages 991-993 RelatedInformation