JP6887599B2 - Equipment and methods for measuring growth or degradation kinetics of colloidal particles - Google Patents

Description

The present invention relates to the measurement and observation of particles in a liquid sample using a microscope equipped with a digital video camera.

Nanoparticles (particles smaller than 1 micron in diameter) are by far the most abundant particle-like entities ubiquitous in the Earth's natural environment and are widely used in many applications associated with human activity. There is. There are many types of naturally occurring nanoparticles and artificial (engineering) nanoparticles. Nanoparticles occur in air, water environment, rainwater, drinking water, biofluids, drugs, drug delivery and therapeutic products, and many industrial products in a wide range. Nanoparticles usually occur in polydisperse aggregates, which are characterized by the simultaneous occurrence of particles of different sizes, also by their diameter greater than 1 micron.

Given the widespread use of nanoparticles, the ability to control and accurately characterize their properties can be beneficial for many applications. Conventional methods for measuring nanoparticle properties may be inaccurate for polydisperse samples with mixed nanoparticle sizes, which are common in many applications. Since the light scattered from all the nanoparticles is measured at the same time, it can be difficult to break down the nanoparticles into their component sizes when different particle sizes are present. Other methods have not been able to take into account the large differences in the intensity of scattered light produced by nanoparticles of different sizes across different nanoparticle sizes. In these techniques, low scattering signals from small nanoparticles may not be detected, or high scattering signals from larger nanoparticles may obscure signals from smaller nanoparticles. .. Also in other methods, the measurements cannot take into account the growth rate or decomposition rate of the particles, so snapshots of the size distribution can become inaccurate after a very short time. possible. As a result of these disadvantages, the concentration of nanoparticles of any given size, and therefore the overall size distribution, can be affected by unknown errors.

These methods of detecting nanoparticles (and larger particles) are commonly referred to as darkfield microscopy. Instruments that perform such analyzes are generally from liquid illumination with a precisely defined narrow optical sheet and nanoparticles (although not always) usually at an angle of 90 degrees to the optical sheet plane. It is equipped with a small cell (eg, a cuvette) that allows observation of scattered light. Note that the observation angle does not have to be 90 degrees, it is important that scattered light is observed. Particles of different sizes can be visualized through a camera that captures the light scattered by the particles, and the images have different sizes and intensities (pixels of varying brightness), depending on the size of the particles.

In Patent Document 1 (“Stramski”), which was filed on June 3, 2015, entitled “NanoPARTICLE ANALYZER”, which is incorporated herein by reference in its entirety, these problems With a single color camera that simultaneously records several different colors of scattered light with several light sources and pixels in a bayer pattern that correspond to the three additive primary colors traditionally used in photography. Was addressed by using. In Stramski's technique, the final image is taken from a single recording device, so images of the same colloidal volume in different colors are recorded within the same area of the recording device or sensor, thereby usually in the camera. Provides pixel numbering for a single origin, which is one of the corners of the sensor. This allows the images to be processed in different colors because the positions of the observed particles are given in the same coordinate system. Unfortunately, Stramski does not describe or disclose any method that takes into account the growth or degradation of particles.

"Multi-camera device for observing microscopic movement and counting particles in colloids, and its calibration (MULTI-CAMERA APPARATUS FOR OBSERVATION OF MICROSCOPIC MOVEMENTS AND COUNTING OF PARTICLES IN COLLOIDS AND ITS 20) Patent Document 2 (“Tatarkiewicz”), filed on February 8, 2014, aligns the calibration mask with images from various light sources for more accurate image processing. By introducing methods and methods that can be used, some of Strumski's shortcomings have been overcome. However, again, Tatarkiewicz does not describe or disclose any method that takes into account the growth or decomposition of particles.

Particle growth / decomposition can be of particular interest in various industries. For example, a pharmaceutical company may want to make sure that its drug decomposes at a certain rate so that it can be used in an effective sustained release mode. Moreover, such degradation can be most therapeutically effective when the particles decompose to the nanoscale and do not recombine into larger particles. Another pharmaceutical company may need to determine the time required to crystallize a new drug based on a protein that can be delivered at a higher dose as large crystals. As such, the methods and devices disclosed in Stramski and Tatarkiewicz can help to take a snapshot of the particle size distribution of the drug, but it does not help to provide a rate of degradation (or vice versa). ..

U.S. Patent Application Publication No. 2015/0346076 U.S. Patent Application Publication No. 2017/0227439

Therefore, what is needed is an improved system that effectively measures the growth / degradation kinetics of colloidal particles.

The following is a simplified summary to provide a basic understanding of some aspects of the claimed subject matter. This summary is not an extensive overview and is not intended to identify the main / decisive elements of the claimed subject matter or to delineate the scope of the claimed subject matter. Its purpose is to present some concepts in a simplified form as a prelude to the more detailed explanation presented later.

The devices, systems, and methods described herein elegantly solve the problems presented above. A system for determining the growth / decomposition rate of colloidal particles has been disclosed, which includes a plurality of light sources and a plurality of sensors. The light source is configured to emit a beam of electromagnetic radiation towards the sample chamber that holds the colloidal particles. The chamber allows a portion of the combined beam to be scattered. The scattered portion of the beam is directed to a sensor that detects electromagnetic radiation. The sensor is connected to a processor that activates the light source and captures the image from the sensor. Multiple images may be acquired at time intervals, and for each image the total light intensity level (the sum of all the intensities registered at all pixels) is calculated and then the maximum intensity in the series. Normalized by level. At each time point, average intensity values from multiple images are acquired. An equation that fits the normalized value over time is then calculated and the gradient is determined from the equation. Also, instead of still images at specific time intervals, a set of short images (ie, moving images) may be acquired at time intervals. The average sum of the intensities for each video and for each time interval is calculated and then normalized by the maximum intensity level in the series.

The processor may also set a measurement window that limits how many images are acquired. The measurement window may be based on the total elapsed time, or the total number of images acquired. It can also be based on the calculated gradient. The processor may further set the maximum image intensity level and adjust the exposure time of the sensor when the total image intensity level exceeds the maximum intensity level.

The device may use a plurality of light sources having multiple wavelengths and a plurality of sensors biased to detect only one of the plurality of wavelengths. The system may use a coupling structure to form a coupled beam, or a uncoupling structure (or beam splitter) before some of the scattered beam reaches the sensor. The plurality of light sources may be a single multi-wavelength light source. The sensor may also be a single sensor capable of detecting multiple wavelengths.

If the calculated gradient is negative, it indicates the decomposition of the colloidal particles, and if it is positive, it indicates the growth of the colloidal particles.
Additional embodiments, alternatives, and variations that will be apparent to those of skill in the art are also disclosed herein and are specifically intended to be included as part of the present invention. The present invention is presented only within the scope of the claims as permitted by the Patent Office in this application or related applications, and the following abstract description of an example does not limit the scope of legal protection. , Uncertain, or otherwise unspecified.

The present invention can be better understood with reference to the following figures. The components in the figure are not necessarily proportional to their actual size and are instead emphasized when the exemplary embodiments of the invention are articulated. In the drawings, similar reference numbers specify corresponding parts at any location in different drawings and / or embodiments. It will be appreciated that certain components and details may not appear in the figures to help explain the invention more clearly.

The figure which shows the system for detecting the electromagnetic radiation from a cuvette using a single wavelength source. The figure which shows the system for detecting the electromagnetic radiation of two wavelengths. The figure which shows the system for detecting the electromagnetic radiation of three wavelengths. A graph showing the scattering coefficient vs. particle diameter. A graph showing the normalized total intensity vs. time of particles in a colloidal solution, showing decomposition. A graph showing the normalized total intensity vs. time of particles in a colloidal solution, showing growth or crystallization. A graph showing the normalized total intensity vs. time of particles in a colloidal solution, showing growth or crystallization and subsequent degradation. A flowchart showing a method for determining the growth / decomposition rate of colloidal particles.

References are made herein to some particular examples of the invention, including any of the best embodiments intended by the inventor for carrying out the invention. Examples of these particular embodiments are shown in the accompanying figures. Although the present invention will be described in connection with these particular embodiments, it will be appreciated that it is not intended to limit the invention to the described or exemplified embodiments. On the contrary, it is intended to include alternatives, modifications, and equivalents that may be included within the gist and scope of the invention as defined by the appended claims.

In the following description, a number of specific details will be described in order to provide a complete understanding of the present invention. Certain exemplary embodiments of the invention may be practiced without some or all of these particular details. In some cases, process operations well known to those of skill in the art are not described in detail so as not to unnecessarily obscure the invention. The various techniques and mechanisms of the invention are sometimes described in the singular for clarity. However, it should be noted that some embodiments include multiple iterations or multiple mechanisms of the technique, unless otherwise stated. Similarly, the various steps of the methods shown and described herein are not necessarily performed in the order shown, or in certain embodiments at all. Therefore, some implementations of the methods described herein may include more or less steps than the steps shown or described. In addition, the techniques and mechanisms of the present invention sometimes describe connections, relationships, or communications between two or more entities. Keep in mind that a connection or relationship between entities does not necessarily mean a direct, unobstructed connection, as various other entities or processes may exist or occur between any two entities. I want to be. As a result, the connections shown do not necessarily mean direct, unobstructed connections, unless otherwise stated.

The following list of exemplary features corresponds to FIGS. 1-4 and is provided for ease of reference, with similar reference numbers being similar anywhere in the specification and in the figure. Specify the feature.

System 10A for detecting electromagnetic radiation from cuvettes using a single wavelength.
System 10B for detecting electromagnetic radiation of multiple wavelengths.
Alternative system 10C for detecting electromagnetic radiation of multiple wavelengths.

Light source 15.
First light source 15A at the first wavelength.
Beam 20 of electromagnetic radiation emitted from a light source. A first beam 20A of electromagnetic radiation at substantially the first wavelength.

A second light source 25 at a second wavelength.
A second beam 30 of electromagnetic radiation at substantially the second wavelength.
A third light source 32 at a third wavelength.

A third beam 34 of electromagnetic radiation at substantially a third wavelength.
Beam coupling structure / dichroic mirror 35.
Second beam coupling structure / dichroic mirror 37.

Combined beam 40.
Optical sheet former 45.
Sample chamber / cuvette 50.

Scattered light 55.
Part of the scattered third beam 55A.
Imaging objective lens 60.

Beam split structure / dichroic mirror 65.
Scattered beam directed at the image sensor.
Separated first wavelength emission 70A.

Image sensor 75.
A first sensor 75A biased to detect electromagnetic radiation at substantially the first wavelength.

Separated second wavelength emission 80.
A second sensor 85 biased to detect electromagnetic radiation at substantially the second wavelength.

Separated third wavelength emission 86.
A third sensor 87 biased to detect electromagnetic radiation at substantially a third wavelength.

Second beam split structure / dichroic mirror 88.
Processor 90.
Best fit line 95 with a constant gradient.

Best fit line 100 with varying gradients.
Method 405 for determining the growth / degradation rate of colloidal particles.
Various steps of the method 410-500.

With reference to FIG. 1A, a system 10A for detecting electromagnetic radiation from a cuvette is shown. System 10A includes a single light source 15 that emits a beam of electromagnetic radiation 20 towards the optical sheet former 45. The resulting optical sheet is directed to a sample chamber / cuvette 50 containing particles, ie, nanoparticles or colloids containing micron-sized particles (not shown). Such cuvettes, which are incorporated herein by reference, are filed June 28, 2016, "SPECIAL PURPOSE CUVETTE for optical microscopy of nanoparticles in liquids. It may be configured in accordance with US Pat. No. 9,541,490, entitled "ASSEMBLY AND METHOD FOR OPTICAL MICROSCOMPY OF NANOPARTICLES IN LIQUIDS".

A part of the optical sheet becomes scattered light 55 when it collides with particles existing in the colloidal solution contained in the cuvette 50, and focuses the imaging objective lens 60 such as a microscope provided with another long working distance objective lens. By matching, it is generally possible to observe at an angle of 90 degrees. Note that the observation angle does not have to be 90 degrees, it is important that scattered light is observed. The scattered light emitted from the imaging objective lens 60 reaches the sensor 75 connected to the processor 90.

1B and 1C show the use of different wavelength and wavelength sensors to reach a more robust system. The advantage of using multiple wavelengths of light is that it expands the range of particle sizes that can be detected. Specifically, the intensity of scattered light depends very strongly on, for example, the particle size, which varies by several orders of magnitude from nanoparticles with a diameter of 10 nm to nanoparticles with a diameter of 1000 nm. A typical sensor assigns each pixel and each color to 8 bits or 256 different values, a zero value corresponds to the absence of registered light, while a maximum value of 255 is for the system. Corresponds to maximum brightness depending on gain and exposure setup. If any pixel receives more light than the maximum level corresponding to the value 255 (saturation), it reduces the gain of the detector or shortens the exposure time, thus reducing all recorded intensities. It is not possible to distinguish and register such values unless by shifting to lower values. Reducing the gain or shortening the exposure can help distinguish the particles that have saturated the sensor, but these adjustments also reduce the sensitivity of the sensor to the bottom of the spectrum, i.e., smaller particles.

Since the typical size of the nanoparticles (diameter less than 1 micron) is comparable to the wavelength of visible light, the system cannot distinguish the details of the light-scattering nanoparticles, which are scattered at each particle and within the sensor. Only the total intensity of light is recorded, projecting an image that looks like a circular stain or disk covering some of the pixels in the light. For one type of particle with one wavelength and a particular index of index, the intensity of visible light scattered on the nanoparticles is shown in FIG. 2 as a function of the diameter of the scattered particles (calculations are: For a polystyrene sphere with a light wavelength of 450 nm and a refractive index of n = 1.6 in water, it was performed using the so-called Mee theory of scattering at an angle of 90 degrees). For particles with a diameter of less than 100 nm, the scattered intensity is very small and difficult to detect.

Moreover, the scattering efficiency of the particles depends on the wavelength of the light it is exposed to, and therefore the range of detection depends on the wavelength. By using multiple light sources with different wavelengths and detecting those wavelengths separately (eg, the three colors red, green and blue as taught in Stramski and Tatarkiewicz), the operator is registered. By covering a wider range of particle sizes, the dynamic range of the system can be substantially expanded.

With reference to FIG. 1B, an exemplary device using multiple wavelengths is shown, as described in Tatarkiewicz, incorporated herein by reference. Such a system 10B may include a first light source 15A at a first wavelength and a second light source 25 at a second wavelength, such as two lasers having different beam colors or wavelengths. It is also possible to have a single light source capable of producing light at multiple wavelengths.

Each of these two beams is a dichroic mirror that combines the beams from light sources 15 and 25 into a single combined beam 40 and directs the combined beam 40 toward an optical system such as an optical sheet former 45. Is directed to the coupling structure 35 such as. The optical sheet former 45 may include a cylindrical lens along with a long working distance objective lens that forms a very narrow sheet of illumination. The light sheet can be directed to a transparent sample chamber 50 (such as a cuvette).

Part 55A of the coupled beam scattered when colliding with the particles present in the colloidal solution contained in the cubet 50 has the same wavelength as the illumination light from the optical sheet former 45 and has another long operation. By focusing the imaging objective lens 60 of a microscope or the like including a distance objective lens, it is generally possible to observe at an angle of 90 degrees. Note that the observation angle does not have to be 90 degrees, it is important that scattered light is observed. The scattered light emitted from the imaging objective lens 60 is at the component wavelength, that is, the separated first wavelength radiation 70A and the separated second wavelength in the beam dividing structure 65 such as the second dichroic mirror. Divided into radiation 80 and they are two sensors 75A, 85 (digital grayscale) tuned to detect electromagnetic radiation at substantially first and second wavelengths 15A, 25, respectively. It can reach the sensor placed inside the camera independently). The two sensors can also be a single sensor capable of detecting electromagnetic radiation at multiple wavelengths.

The system has more wavelengths, and more by adding more pairs of suitable dichroic mirrors 35, 65 to combine and divide more wavelengths of illumination sources 15, 25. It can be easily extended to the corresponding sensors 75A, 85. Such an exemplary system 10C is shown in FIG. 1C, which is a third light source 32 at a third wavelength that substantially produces a third beam 34 of electromagnetic radiation at a third wavelength. And a third wavelength system with a second coupled structure / dichroic mirror 37. On the detection side of system 10A, a second beam split structure / dichroic mirror 88 is biased to detect a third wavelength emission 86, substantially at a third wavelength. The sensor 87 of 3 separates it so that it can be detected. The sensor (75A, 85, 87) may be connected to a processor 90 that processes the image detected by the sensor (75A, 85, 87).

As mentioned earlier, the sensor records the intensity numerically for each pixel and each wavelength, and generally each pixel and each wavelength has an 8-bit number (corresponding to 256 different values). The assigned, zero value corresponds to the absence of registered light, while the highest value of 255 corresponds to the maximum brightness. The final image obtained from the sensor consists of a matrix of stored numbers corresponding to all pixels available on the sensor, typically over one million pixels. By summing all these numbers, the total brightness of the image as a single number (separate for each wavelength when more than one wavelength is used). Of the light scattered by the particles, which is progressively proportional to the number and size of the particles present in the colloid being analyzed, by acquiring the images at a preselected time, usually at regular time intervals. A series of numbers that represent the temporal evolution of intensity.

Such a value-to-time plot after normalization to the initial value, shown in FIG. 3A, is a process that is present in the colloid and experiences degradation or growth due to some chemical or physical process. Allows an estimate of the rate of decomposition or growth (ie, kinetics) for the number and size of particles in a colloid. Line 95 is the best fit for the graph and the slope of the line is negative, indicating decomposition. Also, the gradient of line 95 represents a single average growth / degradation rate over the observation time. The finer line 100 fits the data, it changes its slope over time, for example, the line 100 has a steep slope for the first minute, which becomes less steep in the next few minutes. .. This suggests that the colloidal solution decomposes rapidly for the first minute and not so rapidly thereafter. Sufficient characterization of growth / degradation rates can be extremely useful in industrial applications such as pharmaceuticals. The slope of the degradation or growth curve is usually associated with the so-called order of the process, for example, the linear time dependence of the degradation rate (when the data is plotted as the log-to-time of drug release or crystallization). A primary process, or a process in which the amount of drug applied to a released drug is proportional to the amount of decrease in drug released per unit time.

FIG. 3B shows the normalized intensities of the different colloidal solutions plotted against time, indicating that the particles are growing, that is, crystallizing or agglomerating. .. Line 105 is the best fit for this particular plot, where line 105 has an inflection point in region 110, suggesting that there is a significant amount of crystallization or aggregation during this time. There is. FIG. 3C is another plot of intensity over time for another colloidal solution, which introduces crystallization or aggregation followed by subsequent changes to the colloid (such as the addition of some salt with varying pH). It shows the decomposition after being done.

Here, with reference to FIG. 4, a method 405 for determining the growth / decomposition rate of colloidal particles is described. Although this method is described in steps, it should be noted to those skilled in the art that the sequence of steps can be varied and is still within the scope of the following claims. ..

In step 410, the colloidal solution is inserted into a sample chamber, eg, a cuvette. Steps 415, 420, and 425 set a number of variables for measurement, including the number of images acquired, the time delay between images, and the exposure time. The combination of the delay between images and the number of images acquired defines the measurement window. It can be preset, or it can be dynamic, as described below.

The method may have optional steps 430-460 that address the sensitivity of the system. Specifically, in step 430, the maximum image intensity level is set, in steps 435-445, the light source is activated, the image is captured, and in step 450, the total image intensity level for the image is determined. To do. In step 455, if the total image intensity level from step 450 exceeds the maximum image intensity level, the system reduces the exposure time in step 460 until the total image intensity level falls below the maximum value in steps 435- When 455 is repeated and falls below, in step 465, the system begins to acquire the image and intensity level from which the growth / degradation rate is determined. This helps prevent large particles from supersaturating the image, which tends to prevent the system from seeing smaller particles, which negatively impacts the efficiency and extent of the system.

Steps 430-460 can be omitted as an option and the method can proceed directly from step 425 to step 465, which comprises the first and second light sources (or figures) during the exposure time. If a single light source setup such as that shown in 1A is used, activate (step 465) a single light source and display the first and second images (or in a single light source setup). (Single image) is acquired (steps 468, 470). The system then delays for a preset time period between the images and determines in step 480 whether the number of images has been reached. When the total number of images is reached, the total image intensity level for each image is determined and normalized (steps 485, 490), an expression that fits the normalized values is determined (step 95), and the gradient. Is calculated from the formula (step 500). The normalization process involves finding the maximum intensity (count) of all the pixels in the image and then dividing all the intensities (counts) in this series by that number.

Method 405 can be made more robust by acquiring short moving images instead of a single image in steps 468 and 470. If this is done, in step 486, the average intensity number for each of the videos in the sequence can be calculated for each time interval (ie, the combined intensity of each frame / image in the video is in the video. (Divided by the number of frames / images), then its value is normalized. By performing step 486, method 405 can acquire images or moving images at each time interval and does not unfairly weight one in normalization. Alternatively, if method 405 uses only videos and each video contains the same number of frames / images, then the intensities of all frames / images in each video are used, It may be normalized and therefore step 486 is skipped.

The system does not have to set the total number of images / videos reached (ie, step 415), instead the system can set the total elapsed time and step 480 was the elapsed time met. Note that you can check if. In addition, the processor may determine the total image intensity value and gradient at about the same time as the image was acquired (ie, after step 470). This allows the system to have a dynamic total measurement window. Specifically, if the solution (as in Figure 3A) diminishes at a remarkable rate for the first minute and then stabilizes to be a nearly linear function, the method is to image the image. Approximately at the same time as the acquisition (ie, after step 470), steps 485-500 can be performed and step 480 can be based on an inquiry as to whether the determined gradient is changing. If it has not changed, the method can stop the measurement.

Also note that the delay between images / videos can be dynamic. For example, if the processor determines the total image intensity value and gradient at about the same time that the image is acquired (ie, after step 470), it can determine the gradient at about the same time. If the gradient is larger or changes rapidly (as in the first part of the graph in FIG. 3A), reducing the delay time between subsequent images / videos, in other words, the colloidal solution is rapid. It may be advantageous to obtain more sample images / moving images when changing to. This allows the system to measure gradients more accurately when growth / degradation rates are most variable. The delay between images / moving images can be increased when the speed begins to stabilize and becomes less variable.

Although embodiments herein refer to nanoparticles, the same methods and devices disclosed herein are more than larger particles, such as micron-sized particles and (even over 100 microns). It can also be applied to large particles, and therefore the following claims should not be limited to nanoparticles alone.

Although exemplary embodiments and uses of the invention have been described herein, including those described above and shown in the exemplary diagrams included, the present invention is an exemplary embodiment. There is no intent that the embodiments and uses work, or that exemplary embodiments and uses work, or that they are limited to the methods described herein. In fact, many modifications and modifications to the exemplary embodiments are possible, as will be apparent to those skilled in the art. The method is optional as long as the resulting device, system, or method is within the scope of one of the claims, as permitted by the Patent Office, based on this patent application or any related patent application. Can include devices, structures, methods, or functionality of.

Claims (23)

A system for determining the growth / decomposition rate of colloidal particles.
It comprises a light source configured to emit a sheet of electromagnetic radiation towards the sample chamber, the sheet defining a plane, the chamber holding the colloidal particles, and a portion of the sheet by the colloidal particles. Configured to allow scattering,
The system comprises a sensor provided at a position offset from the plane to observe the scattered portion of the sheet , the sensor being adapted to detect the electromagnetic radiation.
The system further comprises a processor connected to the sensor, which is a processor.
a. The process of activating the light source and
b. A step of acquiring an image from the sensor , wherein the image is composed of an array of pixels, and each pixel is assigned an intensity level representing the intensity of the electromagnetic radiation detected at the position of the pixel .
c. A step of repeating step (b) at intervals of time, and a step of repeating step (b).
d. For each image acquired in step (b), a step of determining the total image intensity level by summing the intensity levels of the pixels in the image.
e. A step of normalizing the total image intensity level for each image determined in step (d), and
f. In step (e), the step of calculating an equation that fits the normalized value, and
g. A system configured to perform the steps of calculating the gradient of the equation in step (f). The system according to claim 1, wherein the image of the step (b) includes a moving image having a plurality of images. The system according to claim 2, wherein the total image intensity level includes a step of determining an average intensity level of the plurality of images in each moving image. The processor
The process of setting the measurement window and
The system according to claim 1, further configured to perform a step of repeating step (c) until the measurement window is reached. The system according to claim 4, wherein the measurement window is based on the total elapsed time or the total number of images acquired. The system according to claim 4, wherein the measurement window is based on the gradient calculated from step (g). The system of claim 1, wherein the time interval is based on the gradient calculated from step (g). The first aspect of claim 1, wherein when the gradient calculated from step (g) is negative, it indicates the decomposition of the colloidal particles, and when it is positive, it indicates the growth of the colloidal particles. System. A system for determining the growth / decomposition rate of colloidal particles.
A first light source configured to emit a first beam of electromagnetic radiation at substantially the first wavelength, and
A second light source configured to emit a second beam of electromagnetic radiation at substantially a second wavelength.
The first and second beams are combined into a combined sheet , the combined sheet is directed to the sample chamber, the sheet defines a plane, and the chamber holds the colloidal particles. , A portion of the bonded sheet is configured to allow it to be scattered by the colloidal particles.
The system comprises a first sensor and a second sensor provided in a position offset from the plane in order to observe a portion of disturbed diverging the sheets the coupling, the sensor of said first , Substantially biased to detect electromagnetic radiation at the first wavelength, and the second sensor biased to substantially detect electromagnetic radiation at the second wavelength.
The system further comprises a processor connected to the first and second sensors.
a. The step of activating the first and second light sources and
b. In the step of acquiring an image from the first and second sensors , the image is composed of an array of pixels, and each pixel has an intensity level representing the intensity of the electromagnetic radiation detected at the position of the pixel. The assigned process and
c. A step of repeating step (b) at intervals of time, and a step of repeating step (b).
d. For each image acquired in step (b), a step of determining the total image intensity level by summing the intensity levels of the pixels in the image.
e. A step of normalizing the total image intensity level for each image determined in step (d), and
f. In step (e), the step of calculating an equation that fits the normalized value, and
g. A system configured to perform the steps of calculating the gradient of the equation in step (f). The system according to claim 9, wherein the image in step (b) includes a moving image having a plurality of images. The system according to claim 10, wherein the total image intensity level includes a step of determining an average intensity level of the plurality of images in each moving image. The processor
The process of setting the measurement window and
The system according to claim 9, further configured to perform a step of repeating step (c) until the measurement window is reached. The system of claim 12, wherein the measurement window is based on the total elapsed time or the total number of images acquired. The system according to claim 12, wherein the measurement window is based on the gradient calculated from step (g). The system of claim 9, wherein the time interval is based on the gradient calculated from step (g). The ninth aspect of claim 9, wherein when the gradient calculated from step (g) is negative, it indicates the decomposition of the colloidal particles, and when it is positive, it indicates the growth of the colloidal particles. System. It further comprises a third light source configured to emit a third beam of electromagnetic radiation at substantially a third wavelength, the third beam being coupled to the coupled sheet.
The scattered portion of the bonded sheet is further directed to a third sensor that is biased to detect electromagnetic radiation at substantially said third wavelength.
The system of claim 9, wherein the processor is connected to the third sensor, and step (b) further comprises the step of acquiring an image from the third sensor. The system according to claim 9, wherein the first and second light sources are a single light source. The system according to claim 9, wherein the first and second sensors are a single sensor. The system of claim 9, wherein the first and second beams are coupled by a coupled structure. 9. The system of claim 9, wherein the scattered portion of the coupled sheet is decoupled by a beam splitter before reaching the first or second sensor. The processor
The process of setting the maximum image intensity level and
9. The system of claim 9, further configured to perform a step of adjusting the exposure time of the sensor when the total image intensity level exceeds the maximum intensity level. A method for determining the growth / decomposition rate of colloidal particles.
a. A step of providing a light source configured to emit a sheet of electromagnetic radiation towards a sample chamber, the sheet defining a plane, the chamber holding the colloidal particles and a portion of the sheet . Is configured to allow it to be scattered by the colloidal particles, the scattered portion of the sheet is directed to a sensor located at a position offset from the plane , the sensor being the electromagnetic. Processes and processes that are adapted to detect radiation,
b. The process of activating the light source and
c. A step of acquiring an image from the sensor , wherein the image is composed of an array of pixels, and each pixel is assigned an intensity level representing the intensity of the electromagnetic radiation detected at the position of the pixel .
d. A step of repeating step (c) with a time interval and a step of repeating step (c).
e. For each image acquired in step (c), a step of determining the total image intensity level by summing the intensity levels of the pixels in the image.
f. A step of normalizing the total image intensity level for each image determined in the step (e), and
g. In step (f), the step of calculating an equation that fits the normalized value, and
h. A method comprising the step of calculating the gradient of the equation in step (g).

Images