Technology

One platform, two powerful imaging modalities. Complete, multi-parameter measurement of single EVs.

Interferometric & Fluorescence Imaging

ExoView allows high-throughput sizing and counting of EVs down to 50nm using label-free interferometric imaging. Combined with multi-color, ultrasensitive fluorescence imaging, biomarkers can be colocalized on single EVs.

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Label Free Key Features

The ExoView platform uses interferometric light scatter imaging to sensitivity size and count single specific EVs down to 50nm. The ultrasensitive technique requires only 35µL of sample volume and can be used with or without purification. These powerful features work in combination with the fluorescence modality of the ExoView platform to provide colocalization information.  

 
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PURIFICATION NOT REQUIRED

Bypass purification biases with single particle counting

EV SIZE

High-resolution sizing of single EVs

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Specificity

Definite characterization via immunoaffinity

EV COUNT

Single EV counting based on affinity

 PURIFICATION NOT REQUIRED

As little as 35 µL of unpurified sample can be measured for specific populations of EVs, eliminating variability and biases introduced through sample purification. Other nanoparticle characterization techniques, such as NTA and TRPS, are unable to discriminate EVs from particles that co-purify, and are subject to errors in data interpretation.  ExoView™ bypasses these challenges by measuring only specific EVs using immunoaffinity.

common issues with purification

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 MEASURING EV SIZE

ExoView™ uses Single Particle Interferometric Reflectance Imaging Sensing (SP-IRIS) to measure the size of single EVs by analyzing the enhanced light scatter signal of antibody-bound EVs to the ExoView chip.  Using this methodology, EVs as small as 50nm can be imaged and measured. The measurement of light intensity means that ExoView benefits from drastically improved peak-to-peak resolution when compared with many other technologies such as NTA and can therefore more accurately resolve distinct vesicle sub-populations within a heterogeneous sample.

 MEASURING EV COUNT

The number of EVs bound to a chip based on immunoaffinity can be counted. The graph shows the linearity response of the ExoView instrument within a dynamic range of XX.

 SPECIFICITY

EV characterization is specific via immunoaffinity capture to the ExoView chip. Control tests are performed to ensure that the binding events measured are specific.

ExoView’s microarray format allows for many probes to be measured simultaneously, dramatically increasing throughput. The ExoView Tetraspanin Kit contains arrays that have been functionalized with antibodies against CD9, CD63, CD81, CD41a plus IgG negative control. Additionally, customers can work with NanoView Biosciences to design arrays with antibodies against an antigen of their choice.

Key Features of Fluorescent Measurements

The ExoView platform uses ultrasensitive multicolor fluorescence to measure and report colocalization of surface and cargo markers on single EVs. These powerful features work in combination with the label-free count and size modality of the ExoView platform.   

 
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multicolor Fluorescence

Automated 3 color fluorescence imaging across the visible spectrum

Biomarker colocalization

Measure EV sub-populations through relative protein expression of multiple markers

 
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EV CARGO

Probe for cargo in single EVs

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Sensitivity

Enhanced chip for ultrasensitive quantitation

MULTICOLOR FLUORESCENCE

 

The ExoView platform will automatically measure and analyze up to 3 channels of stained single EVs using a range of available fluorescent probes (e.g. antibodies) across the visible spectrum.

BIOMARKER COLOCALIZATION

The ExoView™ platform provides the ability to measure up to 4 markers on a single EV. Biomarker colocalization allows unique sub-populations of extracellular vesicles to be identified and measured.  All EVs are simultaneously sized and counted.


Blue Spots are vesicles that are positive for CD41a + CD63.
Green Spots are vesicles that positive for CD41a + CD81.
Red Spots are vesicles that are positive for CD41a + CD9.
Other colors are vesicles that express various combinations of R/G/B fluorescent signal and CD41a (via capture on surface).

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EV CARGO

EVs on the ExoView™ chip can be permeabilized and probed for luminal proteins and other internal cargo. 3 color fluorescence allows staining of 3 surface and/or internal biomarkers on single extracellular vesicles.

These experiments show the ability to detect luminal proteins such as ALIX and Sytenin through the permeabilization of bound EVs.

 SENSITIVITY

The ExoView platform provides single binding event sensitivity, which mean that even poorly expressed biomarkers on the smallest of extracellular vesicles are detectable.