Electromechanical approach for cancer detection

Patent metadata

Field	Value
Publication number	US-10620150-B2
Application number	US-201615238917-A
Country	US
Kind code	B2
Filing date	Aug 17, 2016
Priority date	Dec 5, 2015
Publication date	Apr 14, 2020
Grant date	Apr 14, 2020

Abstract

A single-cell-based electromechanical method for cancerous state detection including the steps of preparing a suspension of individually suspended biological cells, extracting a single cell from the suspension, holding the extracted single cell from the suspension, measuring a first electrical response of the held single cell, step-wised mechanical aspirating the held single cell to form a mechanically deformed cell, and measuring an electrical response of the held single cell after each step of mechanical aspirating. The cancerous state of the single cell is determined based on the changes in the measured electrical responses.

First claim

What is claimed is:

1. A single-cell-based electromechanical method for cancerous state detection, comprising:
preparing a suspension of individually suspended biological cells having elastic cell membranes, the cells comprising at least one of epithelial cells, endothelial cells, and mesenchymal cells;
extracting a single cell from the suspension using an electrically activated micropipette;
holding the extracted single cell using the electrically activated micropipette;
measuring a first electrical response of the held single cell;
mechanically aspirating the held single cell using the electrically activated micropipette to form a mechanically deformed cell, comprising applying stepwise increasing pressure to the held single cell;
measuring an electrical response of the held single cell after each step of mechanical aspiration, wherein the first electrical response and the electrical response measured after each step of mechanical aspiration of the held single cell comprise an electrical impedance of the held single cell, an electrical phase of the held single cell, and combinations thereof; and
determining the cancerous state of the single cell based on the changes in the measured electrical responses, wherein the average impedance change of an aspirated healthy cell is about ten-fold higher than that of a cancerous cell and the average phase change of an aspirated healthy cell is at least two-fold higher than that of a cancerous cell.

2. The method according to claim 1 , wherein preparing the suspension of individually suspended biological cells includes steps of:
culturing a plurality of biological cells on a substrate;
washing the cultured cells;
trypsinizing the washed cells to detach the cultured cells from the substrate to form a solution; and
centrifuging the solution to separate a cell suspension that includes individually suspended biological cells.

3. The method according to claim 2 , wherein culturing the plurality of biological cells on the substrate is done in a cell culture medium comprising a Roswell Park Memorial Institute-1640 (RPMI-1640) medium.

4. The method according to claim 3 , wherein the culture medium is supplemented with 5% Fetal bovine serum (FBS) and 1% of penicillin and/or streptomycin.

5. The method according to claim 2 , wherein:
the cultured cells are washed with a Phosphate-buffered saline (PBS) solution;
and the washed cells are trypsinized with a solution comprising trypsin and ethylenediaminetetraacetic acid (EDTA).

6. The method according to claim 1 , wherein the epithelial cells comprise lung cells, colon cells, breast cells, prostate cells, gastric cells, and combinations thereof.

7. The method according to claim 1 , wherein the electrically activated micropipette includes a glass micropipette coated with an electrical conductive layer.

8. The method according to claim 1 , wherein the electrically activated micropipette has a diameter in a range of 4.5 μm to 5 μm.

9. The method according to claim 1 , wherein:
the electrically activated micropipette includes a glass micropipette coated with a gold (Au) layer; and
the electrically activated micropipette includes two ends, wherein one end is connected to a movable water reservoir of a microinjection microscope and the other end has a nozzle configured for applying stepwise increasing pressure.

10. The method according to claim 9 , wherein the gold layer has a thickness of about 10 nm and the gold layer is coated on the glass micropipette via a sputtering system.

11. The method according to claim 9 , wherein the stepwise mechanical aspiration of the held single cell using the electrically activated micropipette comprises applying stepwise positive pressure by upwardly displacing the movable water reservoir, or applying stepwise negative pressure by downwardly displacing the movable water reservoir.

12. The method according to claim 1 , wherein measuring the electrical response of the held single cell is conducted by contacting an electrical probe to the held single cell, and wherein the electrical probe is connected to a signal controlling system configured to apply an electrical signal to the held single cell in contact with the probe and configured to acquire an electrical response from the held single cell corresponding to the applied electrical signal.

13. The method according to claim 12 , wherein the electrical probe is a SiNT/W electrical probe comprising a tungsten (W) supported silicon nanotube (SiNT).

14. The method according to claim 12 , wherein the signal controlling system comprises:
an AC signal source configured to apply the electrical signal to the electrical probe; and
a data acquisition module configured to acquire the electrical response corresponding to the electrical signal from the electrical probe.

15. The method according to claim 14 , wherein the AC signal source is configured to apply a voltage of about 40 mV with a frequency range between 100 Hz and 100 KHz to the electrical probe.

16. The method according to claim 1 , wherein each electrical impedance is measured in a frequency range between 100 Hz and 100 kHz.

17. The method according to claim 1 , wherein each measuring step comprises:
contacting a sharpened tip of the SiNT/W electrical probe to the held single cell;
applying an electrical voltage of 40 mV with a frequency range between 100 Hz and 100 KHz to the SiNT/W electrical probe using an AC signal source of a signal controlling system; and
acquiring an electrical response corresponding to the applied electrical signal from the SiNT/W electrical probe using a data acquisition module of the signal controlling system.

18. A single-cell-based electromechanical method for cancerous state detection, comprising:
preparing a suspension of individually suspended epithelial cells;
extracting a single cell from the suspension using an electrically activated micropipette with two ends, the electrically activated micropipette comprising a nozzle at one end and connected to a water reservoir of a microinjection microscope at the other end;
holding the extracted single cell using the electrically activated micropipette, wherein the single cell is extracted and held via the nozzle;
stepwise mechanically aspirating the held single cell to form a mechanically stretched cell, comprising applying stepwise increasing suction force to the held single cell via the nozzle by displacing the water reservoir;
measuring an electrical impedance of the held single cell after each aspiration; and
determining the cancerous state of the single cell based on the changes in the measured electrical impedances, the average impedance change of an aspirated healthy cell is approximately ten-fold higher than that of a cancerous cell.

19. The method according to claim 18 , wherein measuring the electrical impedance comprises:
contacting a sharpened tip of the SiNT/W electrical probe to the held single cell;
applying an electrical voltage of 40 mV with a frequency range between 100 Hz and 100 KHz to the SiNT/W electrical probe using an AC signal source of a signal controlling system; and
acquiring an electrical impedance corresponding to the applied electrical signal from the SiNT/W electrical probe using a data acquisition module of the signal controlling system.

Assignees

• Abdolahad Mohammad
• Saeidi Ali
• Gharooni Milad

Inventors

• Abdolahad Mohammad
• Saeidi Ali
• Gharooni Milad

Classifications

• C23C28/322

  only coatings of metal elements only · CPC title

• G01N27/283

  Means for supporting or introducing electrochemical probes · CPC title

• C23C28/321

  with at least one metal alloy layer · CPC title

• C23C14/16

  on metallic substrates or on substrates of boron or silicon · CPC title

• G01N27/3278 Primary

  involving nanosized elements, e.g. nanogaps or nanoparticles (nanopores G01N33/48721; magnetic beads G01N27/745) · CPC title