Technology and Innovations at MBI

“It’s still magic even if you know how it’s done.”Terry Pratchett

Technological advances aid and abet scientific understanding and discoveries. At the Mechanobiology Institute, scientists have created tools to isolate circulating tumour cells ( for certain types of cancers), through a non-invasive method and created nano-bits of different topographies, to grow cells on. Supported bilayers to study interactions of molecules, is developing into a tool for drug optimization studies and specially crafted microscopes are being developed to address specific requirements for imaging cellular and tissue dynamics. These and other interesting developments are a spin-off of the active collaborations and open laboratory concept, which are the foundation stones of the Mechanobiology Institute.

The CTChip®

Principal investigator: Lim Chwee Teck

To be able to harvest rare circulating cancer cells, or other cells, from blood is an essential first step in many biological or biomedical assays designed for disease detection, diagnosis and prognosis. CT Lim’s lab is currently developing label-free cell mechanics-based microfluidic devices to detect and diagnose human diseases through the capture of the targeted diseased cells.

interior-ctLimHThe principle makes use of the fact that circulating tumour cells in peripheral have biomechanical and properties that are significantly different from those of blood cells. The devices are microfluidics-based and that have the advantages of reduced sample volumes, faster processing time, higher sensitivity and spatial resolution, low cost and portability. Using this approach, MBI scientists have been developing microdevices that are efficient and effective in detecting diseased cells. The potential is to create true point-of-care lab-on-a-chip systems. One such biochip, the CTChip®, has recently been commercialized by an NUS startup, Clearbridge Biomedics, co-founded by CT Lim. This biochip is undergoing clinical tests in cancer centres and hospitals, both locally and in the USA, Europe, Japan, Korea, Taiwan and Australia.

The 3-year old startup has just received their series B Funding in March 2013. CT Lim and his startup have also won several awards including the Credit Suisse Technopreneur of the Year Award 2012, Wall Street Journal Asian Innovation Award (Gold) 2012, Asian Entrepreneurship Award (First Prize) 2012, InnovFest Promising Startup Award 2012, President’s Technology Award 2011, Faculty Research Award 2011, IES Prestigious Engineering Achievement Award 2010 and Tan Kah Kee Young Inventor’s Award 2009. top

 

The MembraneChip™

Principal investigator: Jay Groves

On March 5, 2012 SARcode Bioscience, a privately held biopharmaceutical company, announced that it has completed patient enrollment in their Phase 3 clinical study of SAR 1118 ophthalmic solution. SAR 1118 is a first-in-class small-molecule integrin antagonist that inhibits T-cell inflammation by blocking the binding of two key surface proteins (LFA-1 and ICAM-1) that mediate the chronic inflammatory cascade associated with dry eye disease, which is estimated to affect approximately 25 million people in the United States. Dry eye is an inflammatory disease that results from miscommunications between cells at intercellular junctions.

grovesDiseases of intercellular interactions have been notoriously resistant to development of therapeutic drugs.  One key reason is that physical properties of cell surfaces intrinsically integrate classical biochemical interactions with larger scale spatial movements and mechanical forces.  Studies of such phenomenon is the basis of the MBI, and core MBI technology is making important contributions to the development of SAR 1118 in an industry collaboration involving the laboratory of Professor Jay T Groves and his MembraneChipTM technology.

The MembraneChip platform integrates biological cell membranes with silica microchips to create highly controlled interfaces with living cells.  Using this technology, the mechanism of action of SARcode’s new drug has been clarified.  This is of significant importance the humans who will receive the drug because the MembraneChip data correctly reveal the functional drug dose, whereas classical biochemical studies suggest an incorrect dose.  Chip fabrication as well as their usage in a variety of research projects is ongoing with the MBI.  The success of this drug development story is triggering strong demand for the technology, and several new commercial projects are getting underway around the world.  top

 

The MARC Chip

Principal investigators: Evelyn Yim, Michael Sheetz

yimIn collaboration with the Institute of Materials Research and Engineering (IMRE), A*STAR, the Mechanobiology Institute’s Dr. Evelyn Yim and Mike Sheetz developed a customizable topography chip for highly efficient high throughput studies of cell-topography interaction. This topography chip, the Multi-Architecture Chip (MARC), is designed and fabricated, based on an innovative concept, to provide a platform of nanotopographies in an array that eliminates the need to analyze one topography at a time, hence overcoming one of the current hurdles in conventional cell-topography interaction studies.

The chip dimension of 2.2 cm x 2.2 cm can fit within standard cell culture plates or dishes and can hold up to 121 different nanopatterned surfaces of 2x2mm. Each area accommodates enough cells for statistical analysis and the whole chip will fit in standard microscopy specimen holder to allow microscopic inspection of cellular responses.

MARC has already been used to optimize the combination of topographical and biochemical cues for stem cell differentiation to neurons. Thus, the customizable MARC as a topography chip is a novel and unique platform at the nanometer level with a wide range of potential biological applications. top

 

L’Oreal and MBI

Principal investigators: Benoit LadouxLim Chwee Teck

The Mechanobiology Institute and L’Oreal embarked on a 2-year research agreement in November 2011 to characterize the behavior of major skin cell types present in the dermis and the epidermis (keratinocytes and fibroblasts) after they were exposed to mechanical stresses and topographical variations of their environment. top

 

Real-time FLIM and the iPALM system

Principal investigator:  Tony Pakorn Kanchanawong

Along with the Zeiss ELYRA PALM, the Nikon NSTORM, and the Olympus-Roper-MicroLambda ILAS2 TIRF system, the MBI hosts two additional single molecule detection fluorescence microscopes which are not yet commercially available.

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The iPALM: the laser module is on the left, then the feeder and camera box, and on the right the lenses and the stage.

The iPALM (Interference Photoactivation Localization Microscope), uses photoconversion or activation to turn a small subset of fluorophores fluorescent and hence image a sparse distribution of them. As each fluorochrome is spatially separated, it becomes a coherent source, i.e., each emitted photon interferes with itself. This interference is used to use interferometry with three cameras to determine the distance of the fluorophore from the focal plane. Hence, the microscope records interferometric patterns rather than images but due to the sparse signals they look like images which change in intensity. No large scale fringes are visible.

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The iPALM: The laser case (left) is here shown open.

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The feeder module.

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Back view showing the width of the instruments and computer cluster.

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The camera module.

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The stage and lenses (on right).

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The camera mode is used to find interesting areas within a cell cluster and then the cell to conduct Fluorescence Lifetime Imaging (FLIM).

Once a location has been chosen, a 100 picosecond pulsed source is used to excite a large fraction of the fluorophores. The light from the selected spot is collected optically and transferred via fiber into the black box to the left of the microscope (shown above).

Where a single photon sensitive avalanche diode is installed:

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the detector issues a signal when struck by a photon. A wavelength filter assures that it is an emitted fluorescent photon.

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The fast counter unit.

The time difference between the excitation pulse and the emitted photon can be clocked to about 100ps accuracy. Repeating this measurement many, many times and omitting the samples where no emitted photon is found, we are given a good distribution of the half life of the excited state of the fluorophore. If steric hindrance, pH, temperature, Ca+, etc., influence that lifetime, this can be observed with this instrument. And because the instrument is single molecule sensitive, this influence can be seen on the molecular level. top

Learn more about MBI’s Microscopy Core.