scholarly journals Review on Complete Mueller Matrix Optical Scanning Microscopy Imaging

2021 ◽  
Vol 11 (4) ◽  
pp. 1632
Author(s):  
Aymeric Le Gratiet ◽  
Ali Mohebi ◽  
Fabio Callegari ◽  
Paolo Bianchini ◽  
Alberto Diaspro
Keyword(s):  
High Speed ◽  
Polarized Light ◽  
Mueller Matrix ◽  
Scanning Microscopy ◽  
Label Free ◽  
Microscopy Imaging ◽  
Excitation Light ◽  
Optical Scanning ◽  
Polarization Control ◽  
Main Challenge

Optical scanning microscopy techniques based on the polarization control of the light have the capability of providing non invasive label-free contrast. By comparing the polarization states of the excitation light with its transformation after interaction with the sample, the full optical properties can be summarized in a single 4×4 Mueller matrix. The main challenge of such a technique is to encode and decode the polarized light in an optimal way pixel-by-pixel and take into account the polarimetric artifacts from the optical devices composing the instrument in a rigorous calibration step. In this review, we describe the different approaches for implementing such a technique into an optical scanning microscope, that requires a high speed rate polarization control. Thus, we explore the recent advances in term of technology from the industrial to the medical applications.

scholarly journals Polarization Label-Free Microscopy Imaging of Biological Samples by Exploiting the Zeeman Laser Emission

2021 ◽  
Vol 9 ◽  
Author(s):  
Fabio Callegari ◽  
Aymeric Le Gratiet ◽  
Alessandro Zunino ◽  
Ali Mohebi ◽  
Paolo Bianchini ◽  
...  
Keyword(s):  
Biological Samples ◽  
Mueller Matrix ◽  
Scanning Microscopy ◽  
Laser Source ◽  
Orthogonal Polarization ◽  
Interference Signal ◽  
Label Free ◽  
Microscopy Imaging ◽  
Polarization States ◽  
Zeeman Laser

Mueller Matrix Microscopy exploits the generation and the analysis of polarized light to create label-free contrast in biological images. However, when dealing with Optical Scanning Microscopy, it is required a fast generation of the polarization states in order to obtain a good Signal-to-Noise Ratio at the pixel-dwell time rate. In this work, we propose a microscopy system based on a scanning beam architecture that is exploiting the simultaneous emission of orthogonal polarization states from a Zeeman laser to provide Mueller Matrix images. This approach is based on the detection of an interference signal that allows to time-encode polarization states directly from the laser source, without the need for further active components for the management of the polarization states. We provide the theoretical model behind this approach and we apply our new method to the imaging of biological samples. Our Mueller Matrix imaging setup enables high-speed scanning microscopy, while preserving compactness and simplicity of construction. Our findings may lead to more effective dissemination of label-free techniques and their use by biological researchers.

scholarly journals Label-Free Chromatin-DNA Imaging by Circular Polarized Light Scattering Scanning Microscopy

2019 ◽  
Vol 116 (3) ◽  
pp. 499a
Author(s):  
Aymeric Le Gratiet ◽  
Riccardo Marongiu ◽  
Luca Pesce ◽  
Michele Oneto ◽  
Paolo Bianchini ◽  
...  
Keyword(s):  
Light Scattering ◽  
Polarized Light ◽  
Scanning Microscopy ◽  
Label Free ◽  
Dna Imaging ◽  
Circular Polarized Light

Label-Free Biosensor Arrays Based on Silicon Ring Resonators and High-Speed Optical Scanning Instrumentation

2010 ◽  
Vol 16 (3) ◽  
pp. 654-661 ◽  
Author(s):  
Muzammil Iqbal ◽  
Martin A Gleeson ◽  
Bradley Spaugh ◽  
Frank Tybor ◽  
William G Gunn ◽  
...  
Keyword(s):  
High Speed ◽  
Ring Resonators ◽  
Label Free ◽  
Biosensor Arrays ◽  
Optical Scanning

scholarly journals Linearly polarized excitation enhances signals from fluorescent voltage indicators

2021 ◽  
Author(s):  
William Bloxham ◽  
Daan Brinks ◽  
Simon Kheifets ◽  
Adam Ezra Cohen
Keyword(s):  
Mammalian Cells ◽  
High Speed ◽  
Signal To Noise ◽  
Excitation Light ◽  
High Background ◽  
Voltage Imaging ◽  
Polarization Control ◽  
Linearly Polarized ◽  
Membrane Bound ◽  
Voltage Indicator

Voltage imaging in cells requires high-speed recording of small fluorescent signals, often leading to low signal-to-noise ratios. Because voltage indicators are membrane-bound, their orientations are partially constrained by the plane of the membrane. We explored whether tuning the linear polarization of excitation light could enhance voltage indicator fluorescence. We tested a panel of dye and protein-based voltage indicators in mammalian cells. The dye BeRST1 showed a 73% increase in brightness between the least and most favorable polarizations. The protein-based reporter ASAP1 showed a 22% change in brightness, and QuasAr3 showed a 14% change in brightness. In very thin neurites expressing QuasAr3, improvements were anomalously large, with a 170% increase in brightness between polarization parallel vs perpendicular to the dendrite. Signal-to-noise ratios of optically recorded action potentials were increased by up to 50% in neurites expressing QuasAr3. These results demonstrate that polarization control can be a facile means to enhance signals from fluorescent voltage indicators, particularly in thin neurites or in high-background environments.

scholarly journals Multimodal Label Free Stokes/Mueller Matrix and Non Linear Scanning Microscopy

2019 ◽  
Vol 116 (3) ◽  
pp. 279a
Author(s):  
Aymeric Le Gratiet ◽  
Riccardo Marongiu ◽  
Paolo Bianchini ◽  
alberto diaspro
Keyword(s):  
Mueller Matrix ◽  
Scanning Microscopy ◽  
Label Free ◽  
Non Linear ◽  
Linear Scanning

scholarly journals Phasor approach of Mueller matrix optical scanning microscopy for biological tissue imaging

2021 ◽  
Author(s):  
A. Le Gratiet ◽  
L. Lanzano ◽  
A. Bendandi ◽  
R. Marongiu ◽  
P. Bianchini ◽  
...  
Keyword(s):  
Biological Tissue ◽  
Mueller Matrix ◽  
Scanning Microscopy ◽  
Tissue Imaging ◽  
Optical Scanning

Five-Dimensional Microscopy using Widefield-Deconvolution: Practical Considerations and Biological Applications

1996 ◽  
Vol 54 ◽  
pp. 268-269
Author(s):  
W.F. Marshall ◽  
K. Oegema ◽  
J. Nunnari ◽  
A.F. Straight ◽  
D.A. Agard ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
High Speed ◽  
Laser Scanning ◽  
Ccd Camera ◽  
Image Plane ◽  
Time Lapse ◽  
Laser Scanning Confocal Microscopy ◽  
Three Dimensions ◽  
Excitation Light ◽  
Cooled Ccd

The ability to image cells in three dimensions has brought about a revolution in biological microscopy, enabling many questions to be asked which would be inaccessible without this capability. There are currently two major methods of three dimensional microscopy: laser-scanning confocal microscopy and widefield-deconvolution microscopy. The method of widefield-deconvolution uses a cooled CCD to acquire images from a standard widefield microscope, and then computationally removes out of focus blur. Using such a scheme, it is easy to acquire time-lapse 3D images of living cells without killing them, and to do so for multiple wavelengths (using computer-controlled filter wheels). Thus, it is now not only feasible, but routine, to perform five dimensional microscopy (three spatial dimensions, plus time, plus wavelength).Widefield-deconvolution has several advantages over confocal microscopy. The two main advantages are high speed of acquisition (because there is no scanning, a single optical section is acquired at a time by using a cooled CCD camera) and the use of low excitation light levels Excitation intensity can be much lower than in a confocal microscope for three reasons: 1) longer exposures can be taken since the entire 512x512 image plane is acquired in parallel, so that dwell time is not an issue, 2) the higher quantum efficiently of a CCD detect over those typically used in confocal microscopy (although this is expected to change due to advances in confocal detector technology), and 3) because no pinhole is used to reject light, a much larger fraction of the emitted light is collected. Thus we can typically acquire images with thousands of photons per pixel using a mercury lamp, instead of a laser, for illumination. The use of low excitation light is critical for living samples, and also reduces bleaching. The high speed of widefield microscopy is also essential for time-lapse 3D microscopy, since one must acquire images quickly enough to resolve interesting events.

Dual-stage piezo nanopositioner for high-speed ion conductance microscopy imaging

2020 ◽  
Vol 28 (10) ◽  
pp. 2203-2214
Author(s):  
Jian ZHUANG ◽  
◽  
Zhi-wu WANG ◽  
Xiao-bo LIAO ◽  
Keyword(s):  
High Speed ◽  
Microscopy Imaging ◽  
Ion Conductance ◽  
Dual Stage
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