HyperCyt Technology
IntelliCyt Technolgy
Modern flow cytometers are capable of making multiparameter measurements on tens of thousands of cells per second in individual samples. By making many sensitive measurements of each cell, there is an abundance of information that can be rapidly acquired. The limitation for flow cytometers has been the ability to achieve high throughput on a multiple sample or microplate level.
IntelliCyt has achieved high throughput flow cytometry by creating a system that introduces the samples to the flow cytometer rapidly and then analyzes the resulting data quickly and efficiently.
The proprietary HyperCyt® sample handling technology enables commercially available flow cytometers to analyze samples from 96- or 384-well microplates, with extremely fast rates and high data quality. Compatible with most flow cytometers, the HyperCyt system is 30 times faster than conventional tube-based approaches.
Based on patented, high-throughput flow cytometry technologies, the HTFC Screening System is providing researchers with new capabilities. Now they can utilize flow cytometry to achieve more results, faster and more economically than ever before.
The Company’s proprietary software technology utilizes a revolutionary approach to flow cytometry data analysis. Data from an entire plate can be quickly analyzed at once to rapidly achieve results and streamline the decision making process.
IntelliCyt has achieved high throughput flow cytometry by creating a system that introduces the samples to the flow cytometer rapidly and then analyzes the resulting data quickly and efficiently.
The proprietary HyperCyt® sample handling technology enables commercially available flow cytometers to analyze samples from 96- or 384-well microplates, with extremely fast rates and high data quality. Compatible with most flow cytometers, the HyperCyt system is 30 times faster than conventional tube-based approaches.
Based on patented, high-throughput flow cytometry technologies, the HTFC Screening System is providing researchers with new capabilities. Now they can utilize flow cytometry to achieve more results, faster and more economically than ever before.
The Company’s proprietary software technology utilizes a revolutionary approach to flow cytometry data analysis. Data from an entire plate can be quickly analyzed at once to rapidly achieve results and streamline the decision making process.
Flow Cytometry Overview
Flow cytometry is a well established technology that has been widely used in many disciplines for measuring intrinsic properties of cells, such as the size and subcellular structures, and extrinsic properties based upon fluorescent biochemical labels to look at a variety of cellular features such as protein binding, metabolic activities or DNA content.
Cells or other particles in liquid suspension, ranging in sizes from less than 1µm to greater than 25µm, are introduced into a moving focused stream that is interrogated by one or more lasers and the resultant fluorescence or scattered laser light is captured by an optical detector and quantified. Only the fluorescence that is associated with the labeled particles is measured, so that background fluorescence in the fluid goes undetected.
Multiple fluorescence measurements can be made simultaneously on a per cell basis by a series of photomultiplier tubes equipped with fluorochrome specific optical filters that collect the emitted fluorescence and correlate the intensity with the specific particle that passed through the laser beam. Additionally the amount of laser light that is “scattered” by the particle is also measured—giving indications of the relative cell size and membrane integrity when captured in line with the incident light source (forward scattered light – FSC), and indicating the amount of sub-cellular complexity or granularity when captured at a 90° angle (side scatter – SSC).
All of the data collected by the detectors/photomultiplier tubes are correlated with each particle detected and stored in a list mode file of the sampled suspension. Flow cytometry analysis software is used to display results for each measured parameter for each cell or particle and is used to make correlations between any of the fluorescent markers and the cell size, sub-cellular structure and all combinations of the same, allowing the identification of subpopulations within the sample.
This ability to make numerous simultaneous and sensitive measurements in a given sample provides the opportunity to conduct experiments wherein many biomarkers or cellular characteristics can be measured on living cells and provide a more thorough investigation into the effects of chemical or biological agents on cellular function.
One of the most important aspects of flow cytometry is the ability to perform multiple analyses on each cell or particle in a sample, known more commonly as multiplexing. The idea of multiplexing is fairly straightforward wherein different types of particles (e.g. cells or beads) containing multiple fluorescent tags, are analyzed together in the same sample. One important advantage provided by multiplexing is the immediate gain in productivity, where for example, using two or more fluorescent endpoints in one flow assay is equivalent to running two or more separate experiments on a plate reader. To this end, the gains in productivity and information per sample can be exponential.
Cells or other particles in liquid suspension, ranging in sizes from less than 1µm to greater than 25µm, are introduced into a moving focused stream that is interrogated by one or more lasers and the resultant fluorescence or scattered laser light is captured by an optical detector and quantified. Only the fluorescence that is associated with the labeled particles is measured, so that background fluorescence in the fluid goes undetected.
Multiple fluorescence measurements can be made simultaneously on a per cell basis by a series of photomultiplier tubes equipped with fluorochrome specific optical filters that collect the emitted fluorescence and correlate the intensity with the specific particle that passed through the laser beam. Additionally the amount of laser light that is “scattered” by the particle is also measured—giving indications of the relative cell size and membrane integrity when captured in line with the incident light source (forward scattered light – FSC), and indicating the amount of sub-cellular complexity or granularity when captured at a 90° angle (side scatter – SSC).
All of the data collected by the detectors/photomultiplier tubes are correlated with each particle detected and stored in a list mode file of the sampled suspension. Flow cytometry analysis software is used to display results for each measured parameter for each cell or particle and is used to make correlations between any of the fluorescent markers and the cell size, sub-cellular structure and all combinations of the same, allowing the identification of subpopulations within the sample.
This ability to make numerous simultaneous and sensitive measurements in a given sample provides the opportunity to conduct experiments wherein many biomarkers or cellular characteristics can be measured on living cells and provide a more thorough investigation into the effects of chemical or biological agents on cellular function.
One of the most important aspects of flow cytometry is the ability to perform multiple analyses on each cell or particle in a sample, known more commonly as multiplexing. The idea of multiplexing is fairly straightforward wherein different types of particles (e.g. cells or beads) containing multiple fluorescent tags, are analyzed together in the same sample. One important advantage provided by multiplexing is the immediate gain in productivity, where for example, using two or more fluorescent endpoints in one flow assay is equivalent to running two or more separate experiments on a plate reader. To this end, the gains in productivity and information per sample can be exponential.
Multiplexing with Flow Cytometry
A number of multiplexing strategies have been put into practice. A simple example includes combining a fluorescent receptor binding readout with cell counts and a cell viability indicator. A more complex example is the assessment of binding events involving multiple targets on individual cells, with each binding event detected by a different color fluorescent marker. This has been taken to an extreme where, in an immunological proof of concept study, as many as 19 separate parameters were analyzed in a single sample by using various cell-surface markers and intracellular cytokine tags.
Another powerful multiplexing approach is to identify different populations within a sample by prelabeling each population with a different color fluorescent marker and then combining them into a single sample. In fact, there are an increasing number of labs interested in pushing the limits in tagging cells this way, also termed “barcoding,” which aims to uniquely label many populations of cells separately, combine them and then deconvolve each population using analysis software. And finally, while there are many ways to multiplex cell-based assays, there are enormous multiplexing capabilities in using bead-based technologies, which can allow for 100-plex, and more recently 500-plex, assays in a bead based ELISA format.
In addition to gains in productivity, multiplexing can be applied to detailed population analyses on samples containing mixed populations. Using cell type specific fluorescent probes, it is possible to identify the subpopulations within a given sample and then query those subpopulations for further information. To illustrate this point, an assay can be constructed that first identifies actively dividing cells and then measure the expression of a specific surface receptor protein within that subpopulation. In another example lymphocyte subsets can be specifically labeled with fluorescent markers, and then queried for expression of cell signaling proteins. In these examples, not only are there multiple endpoints of information within one sample, but differences within the sample are exposed.
Another powerful multiplexing approach is to identify different populations within a sample by prelabeling each population with a different color fluorescent marker and then combining them into a single sample. In fact, there are an increasing number of labs interested in pushing the limits in tagging cells this way, also termed “barcoding,” which aims to uniquely label many populations of cells separately, combine them and then deconvolve each population using analysis software. And finally, while there are many ways to multiplex cell-based assays, there are enormous multiplexing capabilities in using bead-based technologies, which can allow for 100-plex, and more recently 500-plex, assays in a bead based ELISA format.
In addition to gains in productivity, multiplexing can be applied to detailed population analyses on samples containing mixed populations. Using cell type specific fluorescent probes, it is possible to identify the subpopulations within a given sample and then query those subpopulations for further information. To illustrate this point, an assay can be constructed that first identifies actively dividing cells and then measure the expression of a specific surface receptor protein within that subpopulation. In another example lymphocyte subsets can be specifically labeled with fluorescent markers, and then queried for expression of cell signaling proteins. In these examples, not only are there multiple endpoints of information within one sample, but differences within the sample are exposed.