Method of processing dried samples using digital microfluidic device

• US 10,232,374 B2
• Filed: 04/15/2011
• Issued: 03/19/2019
• Est. Priority Date: 05/05/2010
• Status: Active Grant

• Current Assignee: The Governing Council of the University of Toronto (University of Toronto)
• Original Assignee: Miroculus Inc.
• Inventors: Jebrail, Mais J., Yang, Hao, Wheeler, Aaron R.
• Primary Examiner(s): Hixson, Christopher Adam
• Assistant Examiner(s): Adams, Michelle
• Application Number:
  US13/695,869
• Publication Number:
  US 20130164856A1
• Time in Patent Office: 2,895 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: G01N 2001/288 Filter punches
• CPC Class Codes: G01N 2001/4061 Solvent extraction
• CPC Class Codes: H01J 49/00 Particle spectrometers or separator tubes
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

Methods are provided for the preparation of a sample using a digital microfluidic platform and the optional subsequent mass analysis of an extracted analyte. A sample is dried, optionally on a solid phase support, and contacted with digital microfluidic array. An analyte present within the dried sample is extracted into an extraction solvent by electrically addressing the digital microfluidic array to transport a droplet of extraction solvent to the dried sample spot. The extracted sample may be dried and subsequently processed on the digital microfluidic array for derivatization. The digital microfluidic device may further include an integrated microfluidic channel having an output aperture, and the method may further include contacting a droplet containing extracted analyte with the microfluidic channel and applying a suitable electric field for generating nano-electrospray, thereby enabling the device to be directly interfaced with a mass analysis device.

Claims

1. A method of sample preparation in a two-plate digital microfluidic device comprising an array of digital microfluidic elements that can be electrically actuated, the method comprising the steps of:

placing a solid phase support at a first location so that said solid phase support is sandwiched between an upper plate and a lower plate of said two-plate digital microfluidic device, wherein said solid phase support comprises a dried sample, wherein said first location is dropwise addressable under actuation of said two-plate digital microfluidic device, further wherein a lateral extent of said solid phase support is limited to a portion of the array of digital microfluidic elements to permit a transfer of droplets by said array of digital microfluidic elements;

actuating said two-plate digital microfluidic device to transport a droplet of an extraction solvent from a second location that is dropwise addressable under actuation of said two-plate digital microfluidic device to said first location; and

incubating said droplet of said extraction solvent to extract an analyte from said dried sample into said droplet of said extraction solvent, wherein said dried sample is selected from the group consisting of;

dried whole blood, dried serum, dried plasma, dried urine, dried sputum, and dried cerebral spinal fluid.

2. The method according to claim 1 wherein said solid phase support comprises a porous solid phase.

3. The method according to claim 1 wherein said solid phase support comprises filter paper.

4. The method according to claim 3 further comprising punching said filter paper to a pre-selected size prior to placing said solid phase support at said first location.

5. The method according to claim 1 wherein said solid phase support comprises one of:

cellulose, nitrocellulose, and a porous polymer monolith.

6. The method according to claim 1 wherein a lateral extent of said solid phase support is less than one digital microfluidic element of the array of digital microfluidic elements.

7. The method according to claim 1 wherein placing said solid phase support at said first location comprises the steps of:

contacting said solid phase support with said lower plate; and

installing said upper plate.

8. The method according to claim 1 wherein placing said solid phase support at said first location comprises inserting said solid phase support between said lower plate and said upper plate.

9. The method according to claim 1 further comprising receiving, from a remote collection location, said solid phase support comprising said dried sample.

10. The method according to claim 1 wherein said droplet of said extraction solvent is a first droplet of said extraction solvent, said method further comprising:

placing an additional solid phase support at a third location between said upper plate and said lower plate of said two-plate digital microfluidic device, wherein said additional solid phase support comprises a second dried sample, wherein said third location is dropwise addressable under actuation of said two-plate digital microfluidic device;

actuating said two-plate digital microfluidic device to transport an additional droplet of said extraction solvent to said third location;

incubating said additional droplet of said extraction solvent to extract a second analyte from said second dried sample into said additional droplet of said extraction solvent; and

actuating said two-plate digital microfluidic device to contact said additional droplet of said extraction solvent with said first droplet of said extraction solvent.

11. The method according to claim 1 wherein said extraction solvent comprises a polar solvent.

12. The method according to claim 1 wherein said analyte is selected from the group consisting of an amino acid, fatty acid, acylcarnitine, and organic acid.

13. The method according to claim 1 wherein said analyte is a marker of a metabolic disorder.

14. The method according to claim 1 further comprising:

transporting said droplet of extraction solvent after incubation to extract said analyte to a third location that is addressable by said two-plate digital microfluidic device;

drying said droplet of extraction solvent to form a dried droplet of extraction solvent and extracted analyte;

actuating said two-plate digital microfluidic device to transport a droplet of a derivatization solvent to said third location; and

incubating said droplet of said derivatization solvent with said dried droplet of extraction solvent and extracted analyte to obtain a derivatized analyte that is dissolved or suspended in said derivatization solvent.

15. The method according to claim 14 wherein said microfluidic device further comprises a microfluidic channel and an electrical contact for electrically contacting a fluid in said microfluidic channel, wherein an inlet of said microfluidic channel is interfaced with a gap between said upper and lower plate of said two-plate digital microfluidic device such that said two-plate digital microfluidic device may be actuated to contact a droplet with said microfluidic channel and fill said microfluidic channel, and wherein an outlet of said microfluidic channel provides an opening in an external surface of said microfluidic device,

said method further comprising the steps of:

electrically addressing said two-plate digital microfluidic device to transport said droplet of derivatization solvent comprising said derivatized analyte and to contact said droplet of derivatization solvent with said inlet of said microfluidic channel to fill said microfluidic channel;

positioning said outlet of said microfluidic channel adjacent to an inlet of a mass analysis device; and

generating an electrospray cone, said cone emerging from said microfluidic channel and into said inlet of said mass analysis device, by applying a voltage between said electrical contact and said inlet of said mass analysis device.

16. The method according to claim 15 further comprising the step of performing a mass analysis assay for said analyte.

17. The method according to claim 15 wherein said mass analysis device is a tandem mass spectrometer.

18. The method according to claim 15 wherein said electrical contact is located within an opening in said microfluidic channel.

19. The method according to claim 15 wherein said outlet is located at a corner of said microfluidic device.

20. A method of sample preparation in a two-plate digital microfluidic device comprising an array of digital microfluidic elements that can be electrically actuated, the method comprising the steps of:

placing a solid phase support at a first location so that said solid phase support is sandwiched between an upper plate and a lower plate of said two-plate digital microfluidic device, wherein said solid phase support comprises a dried sample, wherein said first location is dropwise addressable under actuation of said two-plate digital microfluidic device, further wherein a lateral extent of said solid phase support is limited to a portion of the array of digital microfluidic elements to permit a transfer of droplets by said array of digital microfluidic elements, wherein said solid phase support comprises filter paper;

punching said filter paper to a pre-selected size prior to placing said solid phase support at said first location;

actuating said two-plate digital microfluidic device to transport a droplet of an extraction solvent from a second location that is dropwise addressable under actuation of said two-plate digital microfluidic device to said first location; and

incubating said droplet of said extraction solvent to extract an analyte from said dried sample into said droplet of said extraction solvent.