Efficient rejection of scattered light enables deep optical sectioning in turbid media with low-numerical-aperture optics in a dual-axis confocal architecture

2008 ◽  
Vol 13 (3) ◽  
pp. 034020 ◽  
Author(s):  
Jonathan T. C. Liu ◽  
Michael J. Mandella ◽  
James M. Crawford ◽  
Christopher H. Contag ◽  
Thomas D. Wang ◽  
...  
Keyword(s):  
Scattered Light ◽  
Numerical Aperture ◽  
Turbid Media ◽  
Optical Sectioning

scholarly journals Optical Characterization of Homogeneous and Heterogeneous Intralipid-Based Samples

2020 ◽  
Vol 10 (18) ◽  
pp. 6234
Author(s):  
Ines Delfino ◽  
Maria Lepore ◽  
Rosario Esposito
Keyword(s):  
Optical Properties ◽  
Light Scattering ◽  
Scattered Light ◽  
Industrial Applications ◽  
Point Of View ◽  
Turbid Media ◽  
Time Resolved ◽  
Propagation Of Light ◽  
First Time

Different scattering processes take place when photons propagate inside turbid media. Many powerful experimental techniques exploiting these processes have been developed and applied over the years in a large variety of situations from fundamental and applied research to industrial applications. In the present paper, we intend to take advantage of Static Light Scattering (SLS), Dynamic Light Scattering (DLS), and Time-Resolved Transmittance (TRT) for investigating all the different scattering regimes by using scattering suspensions in a very large range of scatterer concentrations. The suspensions were prepared using Intralipid 20%, a material largely employed in studies of the optical properties of turbid media, with concentrations from 10−5% to 50%. By the analysis of the angular and temporal dependence of the scattered light, a more reliable description of the scattering process occurring in these samples can be obtained. TRT measurements allowed us to obtain information on the reduced scattering coefficient, an important parameter largely used in the description of the optical properties of turbid media. TRT was also employed for the detection of inclusions embedded in Intralipid suspensions, by using a properly designed data analysis. The present study allowed us to better elucidate the dependence of scattering properties of Intralipid suspensions in a very large concentration range and the occurrence of the different scattering processes involved in the propagation of light in turbid media for the first time to our knowledge. In so doing, the complementary contribution of SLS, DLS, and TRT in the characterization of turbid media from an optical and structural point of view is strongly evidenced.

High-end spectroscopic diffraction gratings: design and manufacturing

2015 ◽  
Vol 4 (1) ◽  
Author(s):  
Tilman Glaser
Keyword(s):  
Scattered Light ◽  
Numerical Aperture ◽  
Diffraction Gratings ◽  
Light Level ◽  
Stray Light ◽  
Maximum Efficiency ◽  
Minimal Volume ◽  
Theoretical Design ◽  
Design And Manufacturing ◽  
Manufacturing Technologies

AbstractDiffraction gratings are key components for spectroscopic systems. For high-end applications, they have to meet advanced requirements as, e.g., maximum efficiency, lowest possible scattered light level, high numerical aperture, and minimal aberrations. Diffraction gratings are demanded to allow spectrometer designs with highest resolution, a maximal étendue, and minimal stray light, built within a minimal volume. This tutorial is intended to provide an overview of different high-end spectroscopic gratings, their theoretical design and manufacturing technologies.

scholarly journals Confocal imaging capacity on a widefield microscope using a spatial light modulator

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0244034
Author(s):  
Yao L. Wang ◽  
Noa W. F. Grooms ◽  
Sabrina C. Civale ◽  
Samuel H. Chung
Keyword(s):  
Cell Body ◽  
Spatial Light Modulator ◽  
Lower Cost ◽  
Scattered Light ◽  
Image Data ◽  
Confocal Microscope ◽  
Confocal Imaging ◽  
Optical Sectioning ◽  
Light Modulator ◽  
Confocal Images

Confocal microscopes can reject out-of-focus and scattered light; however, widefield microscopes are far more common in biological laboratories due to their accessibility and lower cost. We report confocal imaging capacity on a widefield microscope by adding a spatial light modulator (SLM) and utilizing custom illumination and acquisition methods. We discuss our illumination strategy and compare several procedures for postprocessing the acquired image data. We assessed the performance of this system for rejecting out-of-focus light by comparing images taken at 1.4 NA using our widefield microscope, our SLM-enhanced setup, and a commercial confocal microscope. The optical sectioning capability, assessed on thin fluorescent film, was 0.85 ± 0.04 μm for our SLM-enhanced setup and 0.68 ± 0.04 μm for a confocal microscope, while a widefield microscope exhibited no sectioning capability. We demonstrate our setup by imaging the same set of neurons in C. elegans on widefield, SLM, and confocal microscopes. SLM enhancement greatly reduces background from the cell body, allowing visualization of dim fibers nearby. Our SLM-enhanced setup identified 96% of the dim neuronal fibers seen in confocal images while a widefield microscope only identified 50% of the same fibers. Our microscope add-on represents a very simple (2-component) and inexpensive (<$600) approach to enable widefield microscopes to optically section thick samples.

3D Microscopy using Confocal Microscopy

1996 ◽  
Vol 54 ◽  
pp. 270-271
Author(s):  
Ernst H. K. Stelzer ◽  
Steffen Lindek
Keyword(s):  
Incident Light ◽  
Three Dimensional ◽  
Numerical Aperture ◽  
Biological Research ◽  
Half Maximum ◽  
Optical Sectioning ◽  
Observation Volume ◽  
Spread Function ◽  
Relationship Of ◽  
The Relationship

The importance of confocal fluorescence microscopy in modem biological research results from its optical sectioning capability, which allows the three-dimensional analysis of thick specimens. This property is due to the combination of a point-like light source and a point-like detector, which restrict the illumination and detection volumes, respectively. Only the volume that is illuminated and detected is relevant to the confocal observation volume. The smaller it is, the better is the resolution. The performance of a confocal microscope is thus primarily specified by the spatial extent of the confocal point spread function (PSF). The extent can be estimated, e.g., by the volume enclosed by the isosurface at half maximum of the PSF (VHM – volume at half maximum).The relationship of the parameters that determine the lateral resolution of a microscope has been described by Ernst Abbé. The diameter of a light spot in the focal plane Δx is proportional to the wavelength λ of the incident light and inversely proportional to the numerical aperture of the optical system (N.A. = n, ∙ sin α).

Deep optical sectioning in turbid media with dual-axes confocal microscopy: towards in vivo optical biopsy

2008 ◽  
Author(s):  
Jonathan T. C. Liu ◽  
Michael J. Mandella ◽  
James M. Crawford ◽  
Christopher H. Contag ◽  
Thomas D. Wang ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Turbid Media ◽  
Optical Biopsy ◽  
Optical Sectioning
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