Raman super-multiplexed cell and tissue imaging with tunable carbon atom wires

This technology is a method for rapid and time-resolved optical super-multiplexed imaging of cell and tissue components via 20 distinct Raman frequencies achieved by tunable carbon polyyne materials, termed carbon rainbow (Carbow), for distinct spectral barcoding.

Unmet Need: Rapid super-multiplexed optical imaging of cellular and tissue components on relevant timescales

The need for robust and non-invasive imaging methods to identify and capture various tissue (e.g., proteins) and cellular (e.g., organelles or molecules) level targets is becoming increasingly necessary in disease detection and treatment. Existing imaging techniques include radioscopic labeling, fluorescence tagging, magnetic resonance imaging (MRI), and positron emissions tomography (PET). However, these techniques can be invasive, have low spatial resolution, and more importantly, cannot target a wide variety of molecules in a single assay.

The Technology: Engineered polyyne Raman-active compounds for robust multiplexed imaging (>20 colors) and spectral barcoding

This technology uses tunable carbon atom wires with different functional groups to produce 20 distinct Raman signatures which can be used for super-multiplexed optical imaging applications, termed carbon rainbow, or Carbow. The carbon atom wires consist of alkane-alkyne linkages (polyyne groups) of different lengths which can be easily synthesized. These molecules can be used for rapid cellular and tissue-level microscopy, with resolutions as high as organelle or intracell molecular levels. The narrow resonance of the Raman peaks and the 20 distinct signatures give rise to greater than 60,000 distinct spectral imaging barcodes which can be achieved by encapsulating different Carbow combinations in binding materials. Furthermore, due to the molecular level precision of this method, immunological compounds, such as antigens, proteins, and nucleic acids, can be targeted and imaged with high sensitivity through nanoscale amplification.

This technology has been validated in vitro within HeLa cells, hippocampal neurons, and cerebellar mice tissues.

Applications:

• Detection of diseases and personalized assays
• Robust and non-disruptive monitoring of disease progress
• Cell imaging with organelle and molecule-level precision
• Multi-parameter histological data on short timescales
• Non-toxic immunolabeling and immunohistochemistry
• Functionalization for deep-tissue imaging
• Drug-discovery
• Anti-counterfeiting and document security spectroscopic tags

Advantages:

• Encapsulation of polyyne probes for 60,000 distinct spectral imaging barcodes
• 20 tunable polyyne structures with distinct Raman frequencies
• Rapid multicolor microscopy in live cells at organelle and molecular resolutions
• Improves the number of colors for fluorescence-based detection methods
• High sensitivity and nanoscale amplification for super-multiplexed immuno-imaging
• Can be used to image live cells, tissues and organisms without the need to fix the sample
• Can track the distribution and dynamics of small biomolecules that cannot be studied by fluorescence
• Obviates the need for the genetic modifications required by other techniques
• Does not require radioisotope labels
• Is not prone to bleaching – Raman active tags can be imaged over an extended duration

Lead Inventor:

Wei Min, Ph.D.

Patent Information:

Patent Status

Related Publications:

• Zhao Z, Chen C, Wei S, Xiong H, Hu F, Miao Y, Jin T, Min W. “Ultra-bright Raman dots for multiplexed optical imaging” Nat. comms. 2021 Feb 26;12(1)

• Shi L, Wei M, Miao Y, Qian N, Shi L, Singer RA, Benninger RKP, Min W. “Highly-multiplexed volumetric mapping with Raman dye imaging and tissue clearing” Nat Biotechnol. 2022 Mar;40(3):364-373.

• Chen C, Zhao Z, Qian N, Wei S, Hu F, Min W. Multiplexed live-cell profiling with Raman probes” Nat. Comms. 2021 Jun 7; 12(3405).

• Hu F, Zeng C, Long R, Miao Y, Wei L, Xu Q, Min W. “Supermultiplexed optical imaging and barcoding with engineered polyynes” Nature Methods 2018 Mar 01;15(3): 194-200.

• Wei L, Chen Z, Shi L, Long R, Anzalone AV, Zhang L, Hu F, Yuste R, Cornish VW, Min W. “Super-multiplex vibrational imaging” Nature 2017 Apr 27;544(1): 465-470.

• Wie L, Hu F, Chen Z, Shen Y, Zhang L, Min W. “Live-cell bioorthogonal chemical imaging: stimulated Raman scattering microscopy of vibrational tags” Acc. Chem. Res. 2016 Aug 3;49(8): 1494-1502.

• Hu F, Chen Z, Zhang L, Shen Y, Wie L, Min Win Wie L, Hu F, Chen Z, Shen Y, Zhang L, Min W. “Vibrational imaging of glucose uptake activity in live cells and tissues by stimulated Raman scattering” Angew. Chem. Int. Ed 2015 July 16;54(34): 9821.

• Wei L, Shen Y, Xu F, Hu F, Harrington J, Targoff, K, Min W. “Imaging complex protein metabolism in live organisms by stimulated Raman scattering microscopy with isotope labeling” ACS Chem. Biol. 2015 Mar 20;10(3): 901-908.

• Wei L, Hu F, Shen Y, Chen Z, Yu Y, Lin C, Wang MC, Min W. “Live-cell imaging with alkyne-tagged small biomolecules by stimulated Raman Scattering” Nature Methods 2014 Apr 11;4(1): 410-412.

• Chen Z, Paley D, Wei L, Weisman A, Friesner R, Nuckolls C, Min W. “Multicolor live-cell chemical imaging by isotopically edited alkyne vibrational palette” J. Am. Chem. Soc. 2014 May 21;136(22): 8027-8033.

Tech Ventures Reference:

• IR CU13332, CU13346, CU15116, CU17371, CU18220

• Licensing Contact: Beth Kauderer