Flow cytometry able to follow a protein's travel inside cells for improved cancer therapy

Virginia Polytechnic Institute and State University (Virginia Tech; Blacksburg, VA) researchers have developed a flow cytometry-based technique that detects the subcellular location of a protein, which could allow the scientific and technological communities a simple and improved method for studying effectiveness of therapies for disease, including cancer.

Related: Deep-tissue dysplasia detection with real-time subcellular analysis

Virginia Tech chemical engineer Chang Lu and his colleagues have used a National Science Foundation grant to develop their technique, as they recognize that "simple and accessible detection methods that can rapidly screen a large cell population with the resolution of a single cell inside that population has been seriously lacking," he explains.

If a protein is not located in the right subcellular compartment of a cell, "the result can be diseases ranging from metabolic disorders to cancers," Lu explains. "Modulation of protein transport inside a cell is practiced as an important therapeutic approach for cancer treatment. The subcellular location of a target protein can also serve as a useful read-out for high-content screening of cancer drugs."

Virginia Tech chemical engineer Chang Lu and his graduate student Zhenning Cao developed a unique process that could lead to better drug development
Virginia Tech chemical engineer Chang Lu and his graduate student Zhenning Cao developed a unique process that could lead to better drug development. (Image courtesy of Virginia Tech)

In the human body, proteins move between distinct compartments inside cells, including the plasma membrane, the nucleus, and other membrane-enclosed areas. This movement can be a prerequisite for proteins to carry out their intended functions. These functions might include gene transcription and other molecular regulations.

One current evaluation method of protein movement, fluorescence microscopy, can only analyze a limited number of cells, says Lu. And data collected by a second existing assay called subcellular fractionation only reflects the average properties of the cell populations "without revealing the heterogeneity that is often present among seemingly identical cells," Lu and other research team members say in their paper published in the Royal Society of Chemistry journal Chemical Science.

Lu’s team had made some progress in screening cell populations in the past using an electroporation-based technique, but it did not allow the examination of native proteins and primary cells isolated from animals and from patients. So, their new work uses a method that "incorporates selective chemical release of cytosolic proteins with a standard procedure for fluorescent labeling of the protein to detect the subcellular location of a native protein," Lu said. This simple and unique tweak to the conventional cell staining process allowed them to accurately define the subcellular location of the protein by measuring the amount of the residual protein after release. Using a flow cytometer, the speed of such measurement could reach 10,000 to 100,000 cells/s.

A key ingredient to their process is the use of saponin, a class of amphipathic glycosides. It dissolves cholesterol and permeates the plasma membrane to allow protein release. “Gentle treatment by saponin shows minimal effects on the state of the cell,” Lu adds.

To read the Chemical Science paper, please visit http://dx.doi.org/10.1039/C4SC00578C.

-----

Don't miss Strategies in Biophotonics, a conference and exhibition dedicated to development and commercialization of bio-optics and biophotonics technologies!

Follow us on Twitter, 'like' us on Facebook, and join our group on LinkedIn

Subscribe now to BioOptics World magazine; it's free!

Get All the BioOptics World News Delivered to Your Inbox

Subscribe to BioOptics World Magazine or email newsletter today at no cost and receive the latest news and information.

 Subscribe Now
Related Articles

New bioimaging technique offers clear view of nervous system

Scientists at Ludwig-Maximilians University have developed a technique for turning the body of a deceased rodent entirely transparent, revealing the central nervous system in unprecedented clarity....

Fluorescent jellyfish proteins light up unconventional laser

Safer lasers to map your cells could soon be in the offing -- all thanks to the humble jellyfish. Conventional lasers, like the pointer you might use to entertain your cat, produce light by emittin...

Fluorescence microscopy helps provide new insight into how cancer cells metastasize

By using fluorescence microscopy, scientists have discovered an alternate theory on how some cancer cells metastasize.

In vivo imaging method visualizes bone-resorbing cell function in real time

In vivo imaging can visualize sites where osteoclasts (bone-resorbing cells) were in the process of resorbing bone.

BLOGS

Neuro15 exhibitors meet exacting demands: Part 2

Increasingly, neuroscientists are working with researchers in disciplines such as chemistry and p...

Why be free?

A successful career contributed to keeping OpticalRayTracer—an optical design software program—fr...

LASER Munich 2015 is bio-bent

LASER World of Photonics 2015 included the European Conferences on Biomedical Optics among its si...

White Papers

Understanding Optical Filters

Optical filters can be used to attenuate or enhance an image, transmit or reflect specific wavele...

How can I find the right digital camera for my microscopy application?

Nowadays, image processing is found in a wide range of optical microscopy applications. Examples ...

CONNECT WITH US

            

Twitter- BioOptics World

Copyright © 2007-2016. PennWell Corporation, Tulsa, OK. All Rights Reserved.PRIVACY POLICY | TERMS AND CONDITIONS