NanoSight,
leading
manufacturers of unique nanoparticle characterization technology,
reports on
the work of Professor Hang (Hubert) Yin's group at the University of
Colorado
at Boulder where they apply Nanoparticle Tracking Analysis (NTA) to
characterize biological nanoparticles such as microvesicles.
Professor Hang "Hubert" Yin with two members of his research group, Leslie-Anne Morton and Jonel Saludes, use the NanoSight LM10 system.
The Yin Research Lab is interested in studying at the interface of
chemistry,
Biology and engineering with a particular focus on structure-based drug
design,
cell signaling, biochemistry, biotechnology development and membrane
protein
simulations.
The main research goal of the group is to identify and design
peptides
that
sense membrane curvature to better understand protein/peptide-lipid
interactions and potentially create non-invasive probes to detect
highly curved
extracellular vesicles. Currently, we are studying microvesicles as
potential
biomarkers of tumor progression and cancer metastasis. These
nanoparticles are
shed into bodily fluids targeting other cells in the body and are vital
for
inter-cellular communication.
Their experimental protocol involves lipid vesicle preparation by
pressure-controlled extrusion through different membrane pore sizes.
Different
lipid vesicle sizes are prepared in order to mimic the size range of
the
microvesicles that are shed into the extracellular matrix. Following
vesicle
extrusion, it is important to validate the vesicle size. By using
Nanoparticle
Tracking Analysis (NTA) technology, the results provide an accurate
quantification of different populations of vesicle sizes present in the
sample.
Prior to NTA, the group mostly used dynamic light scattering (DLS)
to
determine
the sizes of our synthetic lipid vesicles. Speaking on their use of
NTA,
Professor Yin says "NTA brought several benefits over existing methods.
The detection ranges from 10 - 2000 nm for vesicle sizes, dimensions
that cover
our liposome size of interest. Flow cytometry has a lower limit
detection of
~200 nm to accurately measure particle sizes so did not reach our lower
requirement while DLS measures the average size of all the particles
present in
the sample rather than accurately distinguish different pools of
vesicle sizes,
often creating a bias towards larger particles."
The group has recently published a paper in the Journal of
Visualized
Experimentation that used the NTA technology entitledConstant
Pressure-controlled
Extrusion Method for the Preparation of Nano-sized Liposomes
(Leslie A. Morton, Jonel P. Saludes, Hang Yin). To learn more, please
refer to
doi: 10.3791/4151.