Integrated semiconductor microreactor for real-time monitoring of biological reactions

• US 7,906,321 B2
• Filed: 12/10/2004
• Issued: 03/15/2011
• Est. Priority Date: 12/12/2003
• Status: Active Grant

• Current Assignee: STMicroelectronics SRL (STMicroelectronics NV)
• Original Assignee: STMicroelectronics SRL (STMicroelectronics NV)
• Inventors: Villa, Flavio, Barlocchi, Gabriele, Mastromatteo, Ubaldo
• Primary Examiner(s): Beisner; William H
• Assistant Examiner(s): Hobbs; Michael
• Application Number:
  US11/009,171
• Publication Number:
  US 20050176037A1
• Time in Patent Office: 2,286 Days
• Field of Search: 422/130, 435/288.7
• US Class Current: 435/288.7
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/0097 Optical sensors
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 21/05 Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/6454 using an integrated detector array

Abstract

An integrated semiconductor chemical microreactor for real-time polymerase chain reaction (PCR) monitoring, has a monolithic body of semiconductor material; a number of buried channels formed in the monolithic body; an inlet trench and an outlet trench for each buried channel; and a monitoring trench for each buried channel, extending between the inlet and outlet trenches thereof from the top surface of the monolithic body to the respective buried channel. Real-time PCR monitoring is carried out by channeling light beams into the buried channels, possibly through one of the inlet or outlet trenches, whereby the light beams impinge on the fluid therein and collecting the emergent light coming out from the monitoring trench.

Claims

1. A microreactor comprising:

a. a semiconductor monolithic body;

b. at least one buried channel in said monolithic body and a plurality of heating elements above said buried channel;

c. said semiconductor monolithic body having openings extending through a top surface of the semiconductor monolithic body into said buried channel to fluidly connect an exterior of said semiconductor monolithic body into said buried channel;

d. said openings comprising an inlet opening and an outlet opening, said inlet and outlet openings at opposite ends of said buried channel, and a plurality of monitoring openings therebetween and configured to allow optical inspection of said buried channel;

e. said buried channel further comprising a sloping wall configured to reflect light entering the inlet or outlet openings into said buried channel;

f. wherein each of said monitoring openings is positioned between a pair of said plurality of heating elements;

g. a first transparent material layer arranged to close said monitoring openings;

h. a second transparent material layer of glass over said first transparent material layer;

i. at least one inlet reservoir formed in said first and second transparent material layers above said inlet opening; and

j. at least one outlet reservoir formed in said first and second transparent material layers above said outlet opening.

2. The microreactor of claim 1, wherein said first transparent material layer is a dry resist layer.

15. The microreactor of claim 1, further comprising insulating layer coating said buried channel, wherein said monitoring opening passes through said insulating layer.

3. A process for manufacturing a microreactor comprising:

a. forming at least one buried channel having a sloping wall in a semiconductor monolithic body;

b. forming openings extending through a top surface of the semiconductor monolithic body into said buried channel to fluidly connect an exterior of said semiconductor monolithic body into said buried channel, so that light entering said openings is reflected by said sloping wall into said buried channel, and wherein said openings are an inlet opening and an outlet opening at opposite ends of the buried channel, and a plurality of monitoring openings therebetween;

c. forming a plurality of heating elements above said buried channel, such that each monitoring opening is between a pair of heating elements;

d. forming first and second transparent material layers arranged to close said monitoring openings;

e. forming at least one inlet reservoir in said first and second transparent material layers above said inlet opening; and

f. forming at least one outlet reservoir in said first and second transparent material layers above said outlet opening.

4. The process of claim 3, wherein forming said openings comprises:

a. forming a resist mask over said buried channel;

b. dry etching said monolithic body.

16. The process of claim 3, comprising simultaneously forming said inlet opening, outlet opening and at least one monitoring opening.

5. A method for real-time monitoring a desired product within a fluid in a microreactor, said microreactor comprising:

a. a monolithic body;

b. at least one buried channel having a sloping wall formed inside said monolithic body and containing a fluid; and

c. an inlet opening and an outlet opening formed at opposite ends of said buried channel and extending through a top surface of said monolithic body into said buried channel to fluidly connect an exterior of said monolithic body into said buried channel;

d. a plurality of monitoring openings extending through a top surface of said monolithic body into said buried channel to fluidly connect an exterior of said monolithic body into said buried channel and configured to allow optical inspection of said fluid;

e. a plurality of heaters formed on the monolithic body, over the buried channel, wherein each of said monitoring openings are positioned between a pair of said plurality of heaters;

f. a first and a second transparent material layer arranged to close said monitoring openings;

g. at least one inlet reservoir formed in said first and second transparent material layers above said inlet opening; and

h. at least one outlet reservoir formed in said first and second transparent material layers above said outlet opening;

said method comprising;

i) channeling a light beam into said buried channel, whereby said light beam reflects off said sloping wall into said buried channel; and

ii) collecting and processing a one or more light beams emerging from said monitoring openings.

6. The method of claim 5, wherein said fluid within said buried channel contains a fluorescent reporter and wherein said monochromatic light beam has a wavelength that causes said fluorescent reporter to fluoresce when it impinges thereon.

7. The method of claim 5, wherein said light beam is a monochromatic light beam.

8. The method of claim 5, wherein channeling a light beam comprises:

a. generating a white light beam; and

b. sending said white light beam to a filter.

9. The method of claim 5, wherein said fluid is a biological fluid, a nucleic acid, or DNA.

10. A system for real-time monitoring a desired product within a fluid, comprising:

a. a microreactor including;

i. a monolithic body;

ii. at least one buried channel formed in said monolithic body and containing said fluid; and

iii. inlet and outlet openings formed at opposite ends of said buried channel and extending through a top surface of said monolithic body into said buried channel to fluidly connect an exterior of said monolithic body into said buried channel;

iv. a plurality of monitoring openings extending through said top surface of said monolithic body into said buried channel to fluidly connect an exterior of said monolithic body into said buried channel;

v. a plurality of heaters formed on the monolithic body, over the buried channel, wherein each of said monitoring openings is positioned between a pair of said heaters;

vi. a first and a second transparent material layer arranged to close said monitoring openings;

vii. at least one inlet reservoir formed in said first and second transparent material layers above said inlet opening; and

viii. at least one outlet reservoir formed in said first and second transparent material layers above said outlet opening;

b. a light source generating a light beam arranged over said inlet or outlet openings;

c. a light path extending between said light source and said buried channel, whereby said light beam is channeled into said buried channel and impinges on said fluid within said buried channel; and

d. a light processing arrangement arranged over said monitoring openings for collecting and processing an emergent light emerging from said monitoring opening.

11. The system of claim 10, wherein said light channeling arrangement comprises a deflecting device for deflecting said light beam from said inlet opening into said buried channel.

12. The system of claim 10, wherein said deflecting device is defined by a sloping wall at an end of said buried channel facing said inlet opening.

13. The system of claim 10, wherein said fluid within said buried channel contains a fluorescent reporter and wherein said monochromatic light beam has a wavelength that causes said fluorescent reporter to fluoresce when it impinges thereon.

14. The method of claim 10, wherein said fluid comprises a biological material or nucleic acid or DNA.

17. A microreactor comprising:

a. a semiconductor monolithic body;

b. at least one buried channel in said monolithic body, said monolithic body defining upper, lower and side walls of the buried channel and said walls defining said buried channel;

c. a plurality of heating elements above said buried channel,

d. said monolithic body having openings extending through a top surface of the monolithic body through said upper wall of said buried channel to fluidly connect an exterior of said monolithic body to an interior of said buried channel;

e. said openings comprising an inlet opening and an outlet opening at opposite ends of said buried channel, and a plurality of monitoring openings therebetween and configured to allow optical inspection of said buried channel;

f. said buried channel further comprising a sloping wall configured to reflect light entering the inlet or outlet openings into said buried channel;

g. each of said plurality of monitoring openings are positioned between a pair of said plurality of heating elements;

h. a first and a second transparent material layer arranged to close said monitoring openings;

i. at least one inlet reservoir formed in said first and second transparent material layers above said inlet opening; and

j. at least one outlet reservoir formed in said first and second transparent material layers above said outlet opening; and

k. a sensing electrode arranged in the outlet reservoir.