Laser speckle contrast imaging (LSCI), also called laser speckle imaging (LSI), is an imaging modality based on the analysis of the blurring effect of the speckle pattern. The operation of LSCI is having a wide-field illumination of a rough surface through a coherent light source. Then using photodetectors such as CCD camera or CMOS sensors imaging the resulting laser speckle pattern caused by the interference of coherent light. [1] [2] In biomedical use, the coherent light is typically in the red or near-infrared region to ensure higher penetration depth. [3] When scattering particles moving during the time, the interference caused by the coherent light will have fluctuations which will lead to the intensity variations detected via the photodetector, and this change of the intensity contain the information of scattering particles' motion. [4] Through image the speckle patterns with finite exposure time, areas with scattering particles will appear blurred.
The first practical application of utilizing speckle pattern reduction to mapping retinal blood flow was reported by Fercher and Briers in 1982. This technology was called single-exposure speckle photography at that time. Due to the lacking of sufficient digital techniques in the 1980s, single-exposure speckle photography has a two-step process which made it not convenient and efficient enough for biomedical research especially in clinical use. With the development of digital techniques, including the CCD cameras, CMOS sensors, and computers, in the 1990s, Briers and Webster successfully improved single-exposure speckle photography. It no longer needed to use photographs to capture images. The improved technology is called laser speckle contrast imaging (LSCI) which can directly measure the contrast of speckle pattern. [2] A typical instrumental setup of laser speckle contrast imaging only contains a laser source, camera, diffuser, lens, and computer. [5] Due to the simple structure of the instrumental setup, LSCI can be integrated into other systems easily. [2]
For a fully developed speckle pattern which formed when the complete coherent and polarized light illuminate a static medium, the contrast (K) range from 0 to 1 is defined by the ratio between the standard deviation and mean intensity: [6] [7]
The intensity distribution of the speckle pattern will be used to compute the contrast value.
Autocorrelation functions of electric field are used to measure the relationship between contrast and the motion of scatterers because the intensity fluctuations are produced by electric field changes of scatterers. E(t) is the electric field over time, E* is the complex conjugate of electric field and is the autocorrelation delay time. [7] [6]
Bandyopadhyay et al. showed that the reduced intensity variances of speckle pattern are related to . Therefore, the contrast can be written as [7] [8] [9]
where T is the exposure time. The normalization constant takes into account the loss of correlation due to the detector pixel size, and depolarization of the light through the medium.
Dynamic scatterers' motion can be classified into two categories, one is the ordered motion and the other one is disordered motion. The ordered motion is the ordered flow of scattered while the disordered motion is caused by the temperature effects. The total dynamic scatterers' motions were thought of as Brownian motion historically, the approximate velocity distribution of Brownian motion can be considered as the Lorentzian profile. However, the ordered motion in dynamic scatterers follows Gaussian distribution. When considering the motion distribution, the contrast equation related to the autocorrelation can be updated. The updated equations are as follows, is the contrast equation function in Lorentzian profile and is the contrast equation function in Gaussian profile. is the decorrelation time. Both equations can be used in contrast measurement, some scientists also use contrast equations with the combination of them. However, what the correct theoretical contrast equation should be is still under investigation. [7] [10] [11]
is the normalization constants that vary in different LSCI systems, the value of it is 1, the most common method to determine the value of it is using the following equation. is account for the instability and maximum contrast of each LSCI system.
Static scatterers are present in the assessed sample, speckle contrast produced by static scatterers remains constant. By adding statics scatterers, the contrast equation can be updated again.
*The above equation did not account for the motion distributions.
P1 and P2 are two constants that range from 0 to 1, they are determined by fitting this equation to the actual experimental data. [7]
The relationship between the velocity of scatterers and decorrelation time is as follows, velocity of scatterers such as the blood flow is proportional to the decorrelation time, is the laser light wavelength. [7] [11]
The method to compute the contrast of speckle patterns can be classified into three categories: s-K (spatial), t-K (temporal), and st-K (Spatio-temporal). To compute the spatial contrast, raw images of laser speckle will be separated into small elements, and each element corresponds to a pixels. The value of is determined by the speckle size. The intensity of all the pixels in each element will be summed and averaged to return a mean intensity value (μ), the final contrast value of this element will be calculated based on the mean intensity and actual intensity of each pixel. To improve the resolution limitation, scientists also compute the temporal contrast of the speckle pattern. The method is the same as how to compute spatial contrast but just in temporal. The combination computation of spatial contrast and temporal contrast is Spatio-temporal contrast processing algorithm and this is the most commonly used one. [7] [11]
Compared with other existing imaging technologies, laser speckle contrast imaging has several obvious advantages. It can uses simple and cost-effective instrument to return excellent spatial and temporal resolution imaging. And due to these strengths, laser speckle contrast imaging has been involved in mapping blood flow for decades. The utilize of LSCI has been extended to many subjects in the biomedical field which include but are not limited to rheumatology, burns, dermatology, neurology, gastrointestinal tract surgery, dentistry, cardiovascular research. [11] LSCI can be adopted into another system easily for clinical full-field monitoring, measuring, and investigating living processes in almost real-time scale. [13] [14]
However, LSCI still has some limitations, it can only be used to mapping relative blood flow instead of measuring the absolute blood flow. [13] Due to the complex vascular anatomy structure, the maximum detection depth of LSCI is limited by 900 micrometers now. [7] [15] The scattering and absorption effect of red blood cell can influence the contrast value. [16] The complex physics of measuring behind this technology made it hard to do quantitative measurements. [7]
The Beer-Lambert law is commonly applied to chemical analysis measurements to determine the concentration of chemical species that absorb light. It is often referred to as Beer's law. In physics, the Bouguer–Lambert law is an empirical law which relates the extinction or attenuation of light to the properties of the material through which the light is travelling. It had its first use in astronomical extinction. The fundamental law of extinction is sometimes called the Beer-Bouguer-Lambert law or the Bouguer-Beer-Lambert law or merely the extinction law. The extinction law is also used in understanding attenuation in physical optics, for photons, neutrons, or rarefied gases. In mathematical physics, this law arises as a solution of the BGK equation.
The laser diode rate equations model the electrical and optical performance of a laser diode. This system of ordinary differential equations relates the number or density of photons and charge carriers (electrons) in the device to the injection current and to device and material parameters such as carrier lifetime, photon lifetime, and the optical gain.
Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the differences in the exponential decay rate of the photon emission of a fluorophore from a sample. It can be used as an imaging technique in confocal microscopy, two-photon excitation microscopy, and multiphoton tomography.
Fluorescence correlation spectroscopy (FCS) is a statistical analysis, via time correlation, of stationary fluctuations of the fluorescence intensity. Its theoretical underpinning originated from L. Onsager's regression hypothesis. The analysis provides kinetic parameters of the physical processes underlying the fluctuations. One of the interesting applications of this is an analysis of the concentration fluctuations of fluorescent particles (molecules) in solution. In this application, the fluorescence emitted from a very tiny space in solution containing a small number of fluorescent particles (molecules) is observed. The fluorescence intensity is fluctuating due to Brownian motion of the particles. In other words, the number of the particles in the sub-space defined by the optical system is randomly changing around the average number. The analysis gives the average number of fluorescent particles and average diffusion time, when the particle is passing through the space. Eventually, both the concentration and size of the particle (molecule) are determined. Both parameters are important in biochemical research, biophysics, and chemistry.
In physics, Larmor precession is the precession of the magnetic moment of an object about an external magnetic field. The phenomenon is conceptually similar to the precession of a tilted classical gyroscope in an external torque-exerting gravitational field. Objects with a magnetic moment also have angular momentum and effective internal electric current proportional to their angular momentum; these include electrons, protons, other fermions, many atomic and nuclear systems, as well as classical macroscopic systems. The external magnetic field exerts a torque on the magnetic moment,
Radiation trapping, imprisonment of resonance radiation, radiative transfer of spectral lines, line transfer or radiation diffusion is a phenomenon in physics whereby radiation may be "trapped" in a system as it is emitted by one atom and absorbed by another.
Dynamic light scattering (DLS) is a technique in physics that can be used to determine the size distribution profile of small particles in suspension or polymers in solution. In the scope of DLS, temporal fluctuations are usually analyzed using the intensity or photon auto-correlation function. In the time domain analysis, the autocorrelation function (ACF) usually decays starting from zero delay time, and faster dynamics due to smaller particles lead to faster decorrelation of scattered intensity trace. It has been shown that the intensity ACF is the Fourier transform of the power spectrum, and therefore the DLS measurements can be equally well performed in the spectral domain. DLS can also be used to probe the behavior of complex fluids such as concentrated polymer solutions.
Resonance fluorescence is the process in which a two-level atom system interacts with the quantum electromagnetic field if the field is driven at a frequency near to the natural frequency of the atom.
Electrophoretic light scattering is based on dynamic light scattering. The frequency shift or phase shift of an incident laser beam depends on the dispersed particles mobility. With dynamic light scattering, Brownian motion causes particle motion. With electrophoretic light scattering, oscillating electric field performs this function.
Laser speckle also known as eye testing using speckle can be employed as a method for conducting a very sensitive eye test.
The Kapitza–Dirac effect is a quantum mechanical effect consisting of the diffraction of matter by a standing wave of light. The effect was first predicted as the diffraction of electrons from a standing wave of light by Paul Dirac and Pyotr Kapitsa in 1933. The effect relies on the wave–particle duality of matter as stated by the de Broglie hypothesis in 1924.
Diffusing-wave spectroscopy (DWS) is an optical technique derived from dynamic light scattering (DLS) that studies the dynamics of scattered light in the limit of strong multiple scattering. It has been widely used in the past to study colloidal suspensions, emulsions, foams, gels, biological media and other forms of soft matter. If carefully calibrated, DWS allows the quantitative measurement of microscopic motion in a soft material, from which the rheological properties of the complex medium can be extracted via the microrheology approach.
Ultrasound-modulated optical tomography (UOT), also known as Acousto-Optic Tomography (AOT), is a hybrid imaging modality that combines light and sound; it is a form of tomography involving ultrasound. It is used in imaging of biological soft tissues and has potential applications for early cancer detection. As a hybrid modality which uses both light and sound, UOT provides some of the best features of both: the use of light provides strong contrast and sensitivity ; these two features are derived from the optical component of UOT. The use of ultrasound allows for high resolution, as well as a high imaging depth. However, the difficulty of tackling the two fundamental problems with UOT have caused UOT to evolve relatively slowly; most work in the field is limited to theoretical simulations or phantom / sample studies.
Laser linewidth is the spectral linewidth of a laser beam.
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Speckle variance optical coherence tomography (SV-OCT) is an imaging algorithm for functional optical imaging. Optical coherence tomography is an imaging modality that uses low-coherence interferometry to obtain high resolution, depth-resolved volumetric images. OCT can be used to capture functional images of blood flow, a technique known as optical coherence tomography angiography (OCT-A). SV-OCT is one method for OCT-A that uses the variance of consecutively acquired images to detect flow at the micron scale. SV-OCT can be used to measure the microvasculature of tissue. In particular, it is useful in ophthalmology for visualizing blood flow in retinal and choroidal regions of the eye, which can provide information on the pathophysiology of diseases.
Diffuse correlation spectroscopy (DCS) is a type of medical imaging and optical technique that utilizes near-infrared light to directly and non-invasively measure tissue blood flow. The imaging modality was created by David Boas and Arjun Yodh in 1995.