Key Features of Fluorescent Measurements:
3 color fluorescence. Single binding event sensitivities mean that even the smallest EVs are detectable
Measure up to 4 proteins per EV. Measure EV sub-populations through relative protein expression
Probe for luminal EV proteins and cargo in single EVs
3 COLOR FLUORESCENCE
Measure 3 color fluorescence on single EVs which are bound to the ExoView™ microarray chip.
Extracellular vesicles are captured from solution without purification and bound to one of 6 different capture antibodies (plus control) on the surface of the ExoView™ chip.
Secondary fluorescent antibodies can be added, and up to 3 colors can be measured simultaneously. The secondary antibodies are common to all 6 capture antibodies.
Single binding event sensitivities mean that even poorly expressed proteins on the smallest of extracellular vesicles are detectable through the high sensitivity of ExoView™.
This image shows vesicles bound to a single anti-CD41a spot on the ExoView™ chip. All vesicles imaged on this spot are therefore positive for CD41a.
Once bound, the vesicles have been stained with a 3 color fluorescent cocktail of antibodies against CD63/CD81/CD9.
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).
All vesicles are simultaneously sized and counted.
Biomarker colocalization (4 markers per EV) allows unique sub-populations of extracellular vesicles to be identified and measured.
DETECTING EV CARGO THROUGH FLUORESCENCE
Once bound to the ExoView™ chip, extracellular vesicles can be permeated and probed for luminal proteins and cargo. 3 color fluorescence allows a combination of staining of 3 surface and/or luminal proteins on single extracellular vesicles.
These experiments show the ability to detect luminal proteins such as ALIX and Sytenin through the permeabilization of bound extracellular vesicles.
Measure up to 4 markers on a single extracellular vesicles.