Electrochemical lipidomics for cancer diagnosis
AssigneeNano Hsegar Sazan Salamat Arya.
InventorsMohammad Abdolahad, Ashkan Zandi
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Abstract
A method for detecting cancerous status of a biological sample. The method includes calculating a charge transfer resistance (Rct) of an electrochemical impedance spectroscopy (EIS) associated with lipid secretion of a biological sample, detecting a cancerous state for the biological sample responsive to the calculated Rct being equal to or more than a threshold value, and detecting a normal state for the biological sample responsive to the calculated Rct being less than the threshold value. The biological sample includes a biological sample suspected to be cancerous.
Claims18
1- A method for detecting cancerous status of a biological sample, comprising:
- growing an array of hydrophobic conductive nanostructures on three-integrated electrodes patterned on a catalyst layer deposited on a substrate, the hydrophobic conductive nanostructures comprising vertically aligned multi-walled carbon nanotubes (VAMWCNTs);
- dropping peripheral aqueous media of a biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes, the biological sample comprising a biological sample suspected to be cancerous;
- recording an electrochemical impedance spectroscopy (EIS) from the dropped peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures at an AC voltage between 5 mV and 10 mV by sweeping a plurality of frequency values between 0.01 Hz and 100 kHz, the EIS comprising a semicircular curve;
- calculating a charge transfer resistance (Rct) of the recorded EIS by measuring a diameter of the semicircular curve;
- and
- detecting a cancerous state for the biological sample responsive to the calculated Rct being equal to or more than a threshold value;
- and
- detecting a normal state for the biological sample responsive to the calculated Rct being less than the threshold value.
2- A method for detecting cancerous status of a biological sample, comprising:
- calculating a charge transfer resistance (Rct) of an electrochemical impedance spectroscopy (EIS) associated with lipid secretion of a biological sample, the biological sample comprising a biological sample suspected to be cancerous;
- detecting a cancerous state for the biological sample responsive to the calculated Rct being equal to or more than a threshold value;
- and
- detecting a normal state for the biological sample responsive to the calculated Rct being less than the threshold value.
3- The method of claim 2, wherein each of the detecting the cancerous state for the biological sample and the detecting the normal state for the biological sample comprises comparing the calculated Rct with the threshold value.
4- The method of claim 2, wherein calculating the Rct of the EIS associated with the lipid secretion of the biological sample comprises:
- dropping peripheral aqueous media of the biological sample on an array of hydrophobic conductive nanostructures grown on three-integrated electrodes of a biosensor, the peripheral aqueous media of the biological sample comprising the lipid secretion of the biological sample;
- recording the EIS of the dropped peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures using an electrochemical analyzer;
- and
- measuring a diameter of a semicircular curve of the recorded EIS.
5- The method of claim 4, wherein dropping the peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor comprises dropping the peripheral aqueous media of at least one of a plurality of biological cells, a plurality of biological cell lines, a part of a tissue obtained through surgery or biopsy, a lipid phase of the biological sample, and combinations thereof, on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor.
6- The method of claim 4, wherein dropping the peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor comprises:
- forming a mixture of the biological sample and a solution of metal ions by mixing the biological sample with the solution of metal ions;
- and
- placing the mixture of the biological sample and the solution of metal ions on the hydrophobic conductive nanostructures array grown on the three-integrated electrodes of the biosensor.
7- The method of claim 4, wherein dropping the peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor comprises dropping an extracted lipid phase of the biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor.
8- The method of claim 7, wherein dropping the peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor comprises:
- extracting a lipid phase from the biological sample;
- forming a mixture of the extracted lipid phase and a solution of metal ions by mixing the extracted lipid phase with the solution of metal ions;
- and
- placing the mixture of the extracted lipid phase and the solution of metal ions on the hydrophobic conductive nanostructures array grown on the three-integrated electrodes of the biosensor.
9- The method of claim 8, wherein extracting the lipid phase from the biological sample comprises:
- forming a two-phase mixture of the biological sample in a solution of chloroform and methanol;
- separating a bottom fraction of the two-phase mixture;
- and
- drying the separated bottom fraction of the two-phase mixture.
10- The method of claim 9, wherein forming the two-phase mixture of the biological sample in the solution of chloroform and methanol comprises:
- culturing the biological sample in a cell culture media;
- and
- adding a solution of chloroform and methanol to the cultured biological sample by adding an equal volume of the solution of chloroform and methanol with a volume ratio of 1:2 (Chloroform:methanol) to the biological sample.
11- The method of claim 9, wherein forming the two-phase mixture of the biological sample in the solution of chloroform and methanol comprises:
- absorbing secretion of the biological sample by keeping the biological sample on a foam for a time period between 5 minutes and 30 minutes;
- forming a mixture of chloroform and the secretion of the biological sample by putting the foam containing the secretion of the biological sample in a chloroform solution inside a shaker for a time period between 5 minutes and 30 minutes;
- removing the foam the mixture of chloroform and the secretion of the biological sample;
- and
- adding methanol to mixture of chloroform and the secretion of the biological sample with a volume ratio of 1:2 (Chloroform:methanol).
12- The method of claim 4, wherein dropping the peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures grown on the three-integrated electrodes of the biosensor comprises dropping the peripheral aqueous media of the biological sample on an array of vertically aligned multi-walled carbon nanotubes (VAMWCNTs) grown on the three-integrated electrodes of the biosensor.
13- The method of claim 12, wherein dropping the peripheral aqueous media of the biological sample on the array of VAMWCNTs grown on the three-integrated electrodes of the biosensor comprises dropping the peripheral aqueous media of the biological sample on the array of VAMWCNTs with a length between 2 μm and 12 μm and a diameter between 20 nm and 75 nm for each VAMWCNT of the array of VAMWCNTs.
14- The method of claim 4, wherein recording the EIS of the dropped peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures using the electrochemical analyzer comprises recording the EIS from the dropped peripheral aqueous media of the biological sample on the array of hydrophobic conductive nanostructures at a AC voltage between 5 mV and 10 mV by sweeping a plurality of frequency values between 0.01 Hz and 100 kHz.
15- The method of claim 4, wherein the electrochemical analyzer comprises a potentiostat.
16- The method of claim 4, further comprising fabricating the biosensor by growing the array of hydrophobic conductive nanostructures on the three-integrated electrodes patterned on a catalyst layer deposited on a substrate.
17- The method of claim 16, wherein:
- the substrate comprises at least one of a glass substrate, a silicon substrate, a ceramic substrate, and combinations thereof, and
- the catalyst layer comprises a layer of at least one of iron, cobalt, nickel, and combinations thereof.
18- The method of claim 16, wherein fabricating the biosensor comprises:
- depositing the catalyst layer on the substrate by thermally growing the catalyst layer on the substrate;
- patterning the three-integrated electrodes on the catalyst layer using photolithography technique, the three-integrated electrodes comprising a working electrode, a counter electrode, and a reference electrode;
- and
- growing the array of hydrophobic conductive nanostructures on the patterned three-integrated electrodes using a direct-current plasma enhanced chemical vapor deposition (DC-PECVD) technique.