scholarly journals Imaging analysis of Bartonella species in the skin using single‐photon and multi‐photon (second harmonic generation) laser scanning microscopy

2020 ◽  
Vol 8 (8) ◽  
pp. 1564-1570 ◽  
Author(s):  
Azar Maluki ◽  
Edward Breitschwerdt ◽  
Lynne Bemis ◽  
Rosalie Greenberg ◽  
Bobak Robert Mozayeni ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Laser Scanning ◽  
Single Photon ◽  
Second Harmonic ◽  
Laser Scanning Microscopy ◽  
Scanning Microscopy ◽  
Imaging Analysis ◽  
Bartonella Species

scholarly journals Advanced Label-Free Laser Scanning Microscopy and Its Biological Imaging Application

2021 ◽  
Vol 11 (3) ◽  
pp. 1002
Author(s):  
Xue Wang ◽  
Xinchao Lu ◽  
Chengjun Huang
Keyword(s):  
Laser Scanning ◽  
Second Harmonic ◽  
Biological Tissues ◽  
Biological Imaging ◽  
Laser Scanning Microscopy ◽  
Scanning Microscopy ◽  
Localized Surface Plasmon ◽  
Label Free ◽  
Coherent Raman ◽  
Surface Plasmon Microscopy

By eliminating the photodamage and photobleaching induced by high intensity laser and fluorescent molecular, the label-free laser scanning microscopy shows powerful capability for imaging and dynamic tracing to biological tissues and cells. In this review, three types of label-free laser scanning microscopies: laser scanning coherent Raman scattering microscopy, second harmonic generation microscopy and scanning localized surface plasmon microscopy are discussed with their fundamentals, features and recent progress. The applications of label-free biological imaging of these laser scanning microscopies are also introduced. Finally, the performance of the microscopies is compared and the limitation and perspectives are summarized.

scholarly journals Assessment of Fluorochromes for Two-Photon Laser Scanning Microscopy of Biofilms

2002 ◽  
Vol 68 (2) ◽  
pp. 901-909 ◽  
Author(s):  
Thomas R. Neu ◽  
Ute Kuhlicke ◽  
John R. Lawrence
Keyword(s):  
Nucleic Acid ◽  
Laser Scanning ◽  
Single Photon ◽  
Selective Excitation ◽  
Laser Scanning Microscopy ◽  
Scanning Microscopy ◽  
Excitation Wavelength ◽  
Laser Microscopy ◽  
Two Photon ◽  
Photon Excitation

ABSTRACT A major limitation for the use of two-proton laser scanning microscopy (2P-LSM) in biofilm and other studies is the lack of a thorough understanding of the excitation-emission responses of potential fluorochromes. In order to use 2P-LSM, the utility of various fluorochromes and probes specific for a range of biofilm constituents must be evaluated. The fluorochromes tested in this study included classical nucleic acid-specific stains, such as acridine orange (AO) and 4",6"-diamidino-2-phenylindole (DAPI), as well as recently developed stains. In addition, stains specific for biofilm extracellular polymeric substances (EPS matrix components) were tested. Two-photon excitation with a Ti/Sapphire laser was carried out at wavelengths from 760 to 900 nm in 10-nm steps. It was found that autofluorescence of phototrophic organisms (cyanobacteria and green algae) resulted in strong signals for the entire excitation range. In addition, the coenzyme F420-related autofluorescence of methanogenic bacteria could be used to obtain images of dense aggregates (excitation wavelength, 780 nm). The intensities of the emission signals for the nucleic acid-specific fluorochromes varied. For example, the intensities were similar for excitation wavelengths ranging from 780 to 900 nm for AO but were higher for a narrower range, 780 to 810 nm, for DAPI. In selective excitation, fading, multiple staining, and combined single-photon-two-photon studies, the recently developed nucleic acid-specific fluorochromes proved to be more suitable regardless of whether they are intended for living or fixed samples. Probes specific for proteins and glycoconjugates allowed two-photon imaging of polymeric biofilm constituents. Selective excitation-emission was observed for Calcofluor White M2R (780 to 800 nm) and SyproOrange (880 to 900 nm). In addition, fluor-conjugated concanavalin A lectins were examined and provided acceptable two-photon emission signals at wavelengths ranging from 780 to 800 nm. Finally, CellTracker, a fluorochrome suitable for long-term labeling of microbial eucaryote cells, was found to give strong emission at wavelengths ranging from 770 to 810 nm. If fluorochromes have the same two-photon excitation cross section, they are suitable for multiple staining and multichannel recording. Generally, if an appropriate excitation wavelength and fluorochrome were used, it was possible to obtain more highly resolved images for thick biofilm samples with two-photon laser microscopy than with conventional single-photon laser microscopy. Due to its potential for higher resolution in light-scattering tissue-like material, such as biofilms, and extremely localized excitation, 2P-LSM is a valuable addition to conventional confocal laser scanning microscopy with single-photon excitation. However, further development of the method and basic research are necessary to take full advantage of nonlinear excitation in studies of interfacial microbial ecology.

scholarly journals Second-harmonic generation imaging analysis can help distinguish sarcoidosis from tuberculoid leprosy

2018 ◽  
Vol 23 (12) ◽  
pp. 1
Author(s):  
Fabiane Leonel Utino ◽  
Marina Garcia ◽  
Paulo Eduardo Neves Ferreira Velho ◽  
Andréa Fernandes Eloy da Costa França ◽  
Rafael Fantelli Stelini ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Second Harmonic ◽  
Imaging Analysis ◽  
Tuberculoid Leprosy ◽  
Second Harmonic Generation Imaging

scholarly journals Single-photon-counting detector for increased sensitivity in two-photon laser scanning microscopy

2008 ◽  
Vol 33 (24) ◽  
pp. 2895 ◽  
Author(s):  
Richard K. P. Benninger ◽  
William J. Ashby ◽  
Elisabeth A. Ring ◽  
David W. Piston
Keyword(s):  
Laser Scanning ◽  
Single Photon ◽  
Photon Counting ◽  
Laser Scanning Microscopy ◽  
Scanning Microscopy ◽  
Single Photon Counting ◽  
Photon Counting Detector ◽  
Two Photon ◽  
Increased Sensitivity

scholarly journals Cardiovascular Imaging Using Two-Photon Microscopy

2008 ◽  
Vol 14 (6) ◽  
pp. 492-506 ◽  
Author(s):  
John A. Scherschel ◽  
Michael Rubart
Keyword(s):  
Laser Scanning ◽  
Single Photon ◽  
Second Harmonic ◽  
Multiphoton Microscopy ◽  
Cardiovascular Imaging ◽  
Laser Scanning Microscopy ◽  
Dye Transfer ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

AbstractTwo-photon excitation microscopy has become the standard technique for high resolution deep tissue and intravital imaging. It provides intrinsic three-dimensional resolution in combination with increased penetration depth compared to single-photon confocal microscopy. This article will describe the basic physical principles of two-photon excitation and will review its multiple applications to cardiovascular imaging, including second harmonic generation and fluorescence laser scanning microscopy. In particular, the capability and limitations of multiphoton microscopy to assess functional heterogeneity on a cellular scale deep within intact, Langendorff-perfused hearts are demonstrated. It will also discuss the use of two-photon excitation-induced release of caged compounds for the study of intracellular calcium signaling and intercellular dye transfer.

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