U.S. patent application number 13/478478 was filed with the patent office on 2013-03-07 for optical sensing systems and methods.
This patent application is currently assigned to MEDTRONIC MINIMED, INC.. The applicant listed for this patent is SOREN AASMUL, Henning Munk Ejlersen, Jesper Svenning Kristensen. Invention is credited to SOREN AASMUL, Henning Munk Ejlersen, Jesper Svenning Kristensen.
Application Number | 20130060106 13/478478 |
Document ID | / |
Family ID | 47753655 |
Filed Date | 2013-03-07 |
United States Patent
Application |
20130060106 |
Kind Code |
A1 |
AASMUL; SOREN ; et
al. |
March 7, 2013 |
OPTICAL SENSING SYSTEMS AND METHODS
Abstract
An optical glucose sensor may include an optical fiber and a
glucose-permeable membrane having a hollow interior and being
coupled to the optical fiber's distal end. The membrane's hollow
interior provides a compartment to house a competitive glucose
binding affinity assay. The assay may include a glucose analog that
may be labeled with a dye, and a glucose receptor that may be
labeled with a fluorophore. The optical fiber may include a
compound parabolic concentrator tip, and the compartment may
additionally house a reflector disposed so as to face the optical
fiber's tip. A fluorophore-labeled assay may be interrogated by an
optical interrogating system including a light source and a filter
substrate having one or more coatings to effect, e.g., an
excitation filter and/or an emission filter. The interrogating
system may be manufactured as a stacked planar integrated optical
system and diced into smaller units.
Inventors: |
AASMUL; SOREN; (Holte,
DK) ; Kristensen; Jesper Svenning; (Virum, DK)
; Ejlersen; Henning Munk; (Vedbaek, DK) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AASMUL; SOREN
Kristensen; Jesper Svenning
Ejlersen; Henning Munk |
Holte
Virum
Vedbaek |
|
DK
DK
DK |
|
|
Assignee: |
MEDTRONIC MINIMED, INC.
Northridge
CA
|
Family ID: |
47753655 |
Appl. No.: |
13/478478 |
Filed: |
May 23, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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61531449 |
Sep 6, 2011 |
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61531451 |
Sep 6, 2011 |
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61531456 |
Sep 6, 2011 |
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61554057 |
Nov 1, 2011 |
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61561146 |
Nov 17, 2011 |
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61587819 |
Jan 18, 2012 |
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61620563 |
Apr 5, 2012 |
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Current U.S.
Class: |
600/316 ;
422/69 |
Current CPC
Class: |
G01N 21/7703 20130101;
A61B 5/1459 20130101; A61M 5/1723 20130101; A61B 5/14532 20130101;
A61B 5/0002 20130101; G01N 2021/7786 20130101; G01N 2021/7733
20130101; A61B 5/4839 20130101; A61B 5/14556 20130101; A61B 5/6848
20130101; G01N 2021/772 20130101; G01N 2021/7759 20130101; A61B
5/1473 20130101; A61B 5/6849 20130101; G01N 21/6408 20130101; A61B
5/742 20130101; A61B 5/0004 20130101; G01N 21/6428 20130101; G01N
2021/6441 20130101; G01N 2021/775 20130101; G01N 2333/4724
20130101; G01N 33/66 20130101; A61M 2230/201 20130101; G01N
2021/6484 20130101 |
Class at
Publication: |
600/316 ;
422/69 |
International
Class: |
A61B 5/1459 20060101
A61B005/1459; G01N 30/00 20060101 G01N030/00; A61B 5/1455 20060101
A61B005/1455 |
Claims
1. An optical glucose sensor comprising: an optical fiber having a
proximal end and an opposing distal end; a glucose-permeable
membrane having a hollow interior, an open proximal end, and a
closed distal end, wherein the membrane's proximal end is coupled
to the optical fiber's distal end so as to define a compartment in
said hollow interior between the optical fiber's distal end and the
membrane's distal end; and a competitive glucose binding affinity
assay disposed in said compartment, the assay including a glucose
receptor and a glucose analog.
2. The optical glucose sensor of claim 1, wherein the compartment
is placed within a user's tissue.
3. The optical glucose sensor of claim 2, wherein said proximal end
of the optical fiber is external to the user's body.
4. The optical glucose sensor of claim 3, wherein the proximal end
of the optical fiber is optically coupled to an assay interrogating
system.
5. The optical glucose sensor of claim 4, wherein the interrogating
system is a stacked planar integrated optical system.
6. The optical glucose sensor of claim 2, wherein the proximal end
of the optical fiber is optically coupled to an assay interrogating
system.
7. The optical glucose sensor of claim 6, wherein the interrogating
system is an optical interrogating system.
8. The optical glucose sensor of claim 1, wherein said assay is a
fluorophore labeled assay.
9. The optical glucose sensor of claim 1, wherein said glucose
receptor is labeled with a first fluorophore.
10. The optical glucose sensor of claim 9, wherein the assay
further includes a reference fluorophore different from said first
fluorophore.
11. The optical glucose sensor of claim 10, wherein said first
fluorophore and said reference fluorophore have either different
absorption spectra, or different emission spectra, or both.
12. The optical glucose sensor of claim 10, wherein said first
fluorophore is Alexa Fluor 594 (AF594).
13. The optical glucose sensor of claim 10, wherein said reference
fluorophore is Alexa Fluor 700 (AF700).
14. The optical glucose sensor of claim 10, wherein said reference
fluorophore is labeled onto a macro molecule.
15. The optical glucose sensor of claim 1, wherein said glucose
receptor is selected from the group consisting of Concanavalin A,
glucose galactose binding protein, an antibody, Boronic Acid, and
Mannan Binding Lectin (MBL).
16. The optical glucose sensor of claim 1, wherein the glucose
analog is dextran.
17. The optical glucose sensor of claim 1, wherein the glucose
analog is labeled with a dye.
18. The optical glucose sensor of claim 1, wherein the glucose
receptor is labeled with a fluorophore, the glucose analog is
labeled with a dye, and said fluorophore and dye form a Forster
Resonance Energy Transfer pair.
19. The optical glucose sensor of claim 1, wherein the glucose
receptor is Mannan Binding Lectin (MBL), and the glucose analog is
dextran.
20. The optical glucose sensor of claim 19, wherein the Mannan
Binding Lectin (MBL) is labeled with an Alexa Fluor
fluorophore.
21. The optical glucose sensor of claim 20, wherein the fluorophore
is water soluble.
22. The optical glucose sensor of claim 20, wherein the fluorophore
is Alexa Fluor 594 (AF594).
23. The optical glucose sensor of claim 19, wherein the dextran is
labeled with a dye.
24. The optical glucose sensor of claim 23, wherein the dye is
water soluble.
25. The optical glucose sensor of claim 23, wherein the dye is
hexamethoxy crystalviolet-1 (HMCV1).
26. The optical glucose sensor of claim 1, wherein the glucose
receptor is Mannan Binding Lectin (MBL) labeled with Alexa Fluor
594 (AF594), and the glucose analog is dextran labeled with
hexamethoxy crystalviolet-1 (HMCV1).
27. The optical glucose sensor of claim 26, wherein the assay
further includes a macro molecule labeled with Alexa Fluor 700
(AF700) as a reference fluorophore.
28. The optical glucose sensor of claim 1, wherein said membrane is
tube-shaped.
29. The optical glucose sensor of claim 1, wherein said membrane
comprises a biocompatible polymer.
30. The optical glucose sensor of claim 29, wherein said polymer is
biodegradable.
31. The optical glucose sensor of claim 1, wherein the membrane's
open proximal end is sealably fitted over the optical fiber's
distal end.
32. The optical glucose sensor of claim 1, wherein the distal end
of the optical fiber is straight cut and polished.
33. The optical glucose sensor of claim 1, wherein the distal end
of the optical fiber is in direct contact with the assay.
34. The optical glucose sensor of claim 1, wherein the distal end
of the optical fiber is in the shape of a compound parabolic
concentrator.
35. The optical glucose sensor of claim 34, wherein said compound
parabolic concentrator extends into said compartment and is in
direct contact with the assay in the compartment.
36. The optical glucose sensor of claim 34, further including a
reflector disposed longitudinally spaced apart from a distal end of
the compound parabolic concentrator.
37. The optical glucose sensor of claim 36, wherein the reflector
reflects fluorescence emitted from the assay towards the distal end
of the optical fiber.
38. The optical glucose sensor of claim 36, wherein the reflector
is a concave mirror.
39. The optical glucose sensor of claim 36, wherein the assay is
disposed in a space between the reflector and the distal end of the
compound parabolic concentrator.
40. The optical glucose sensor of claim 34, wherein the compound
parabolic concentrator has a rectangular cross-section.
41. The optical glucose sensor of claim 34, wherein the optical
fiber and the compound parabolic concentrator have circular
cross-sections.
42. The optical glucose sensor of claim 1, wherein the sensor is
implanted within a user's body.
43. The optical glucose sensor of claim 42, wherein the proximal
end of the optical fiber is optically coupled to an assay
interrogating system.
44. The optical glucose sensor of claim 43, wherein the
interrogating system is an optical interrogating system.
45. The optical glucose sensor of claim 44, wherein the
interrogating system is a stacked planar integrated optical
system.
46. The optical glucose sensor of claim 1, wherein the sensor is
biodegradable.
Description
RELATED APPLICATION DATA
[0001] This application claims priority from U.S. Provisional
Application Ser. No. 61/531,449, filed Sep. 6, 2011, and U.S.
Provisional Application Ser. No. 61/531,451, filed Sep. 6, 2011,
and U.S. Provisional Application Ser. No. 61/531,456, filed Sep. 6,
2011, and U.S. Provisional Application Ser. No. 61/554,057, filed
Nov. 1, 2011, and U.S. Provisional Application Ser. No. 61/561,146,
filed Nov. 17, 2011, and U.S. Provisional Application Ser. No.
61/587,819, filed Jan. 18, 2012, and U.S. Provisional Application
Ser. No. 61/620,563, filed Apr. 5, 2012, and is related to the U.S.
Patent Application entitled "Orthogonally Redundant Sensor Systems
and Methods", Attorney Docket No. 040088-0405699, which is being
filed concurrently herewith, all of which are incorporated herein
by reference in their entirety.
FIELD OF THE INVENTION
[0002] Embodiments of the present invention relate generally to
sensor technology, including sensors used for sensing a variety of
physiological parameters, e.g., glucose concentration. More
particularly, embodiments of the invention relate to optical
sensors, to methods of making and using such sensors, to optical
and optoelectronic systems for interrogating optical sensors, and
to methods of making and using such optical/optoelectronic systems.
More particularly still, embodiments of the invention relate to
optical fiber sensors including a fluorophore-labeled assay, to
stacked planar optical integrated systems for interrogating such
optical fiber sensors, and to methods of making and using such
optical fiber sensors and optical integrated systems.
BACKGROUND
[0003] Epifluorescence microscopy is a method of fluorescence
microscopy that is becoming increasingly used in the biological and
medical fields. An epifluorescence microscope is used primarily to
excite a specimen by passing a source light through an objective
lens and then onto the specimen. The fluorescence in the specimen
generates emitted (fluorescent) light which is focused onto a
detector by the same objective lens that is used for the
excitation. Since most of the source light is generally transmitted
through the specimen, only reflected source light reaches the
objective lens together with the fluorescent light. An additional
filter between the objective lens and the detector can filter out
the remaining source light from fluorescent light.
[0004] The underlying principles of epifluorescence microscopy may
be used in optical, or optoelectrical, systems for interrogating
assay-based glucose sensors. The assay in such sensors may be
interrogated using a variety of methods, such as Streak Camera
recording, single photon counting, frequency domain lifetime
measurement, and steady state fluorescence measurement. In both the
frequency domain lifetime and steady state fluorescence
interrogation, the function of the optical interrogation system is
to excite the assay fluorophore(s) and prevent the excitation light
from reaching the detector(s) while, at the same time, transmitting
the emitted fluorescence. It is understood that the fluorescence
emitted from fluorophore-labeled assays is generally weak.
Therefore, it is important to excite the assay as efficiently as
possible and to gather as much of the isotropically emitted
fluorescence as possible.
[0005] In the context of a continuous glucose monitor based on
frequency domain lifetime interrogation and steady state
fluorescence interrogation, it is important not only to minimize
the cost, size, and weight of the (optical system) instrumentation
and of the optical sensor, but also to optimize manufacturability
of both the instrumentation and the sensor. In this regard, the
currently-used optical systems are in general fairly large and
expensive, and require precision assembly as they include a number
of different optical components. Thus, improved optical systems and
optical glucose sensors, including sensors for use with such
optical systems, are needed that address the above-mentioned
requirements.
SUMMARY
[0006] In accordance with one embodiment of the invention, an
optical glucose sensor includes an optical fiber with a
glucose-permeable membrane joined to its distal end. The membrane
may be, e.g., tube-shaped, such that its hollow interior defines a
compartment for holding an assay. In one aspect of the invention,
the assay is a competitive glucose binding affinity assay that
includes a glucose receptor, a glucose analog, a first (donor)
fluorophore labeled onto the glucose receptor, and an acceptor dye
labeled onto the glucose analog. In a variation of this aspect of
the invention, the assay may include a reference fluorophore in
addition to the first fluorophore.
[0007] In accordance with another embodiment of the invention, the
optical fiber of the optical glucose sensor includes a compound
parabolic concentrator (CPC)-shaped tip that is in direct contact
with the assay. In yet another aspect, a reflector may be disposed
within the compartment, opposite the CPC-shaped tip, to reflect
fluorescence that is emitted from the assay towards the CPC-shaped
tip.
[0008] Embodiments of the invention are also directed to optical
systems for lifetime and/or intensity interrogation of the assay.
Thus, in one aspect, a fluorophore-labeled assay may be
interrogated by an optical interrogating system including a light
source and a filter substrate having one or more coatings to
effect, e.g., an excitation filter and/or an emission filter. In
another aspect, the interrogating system may be manufactured as a
wafer-scale stacked planar integrated optical system (SPIOS) and
diced into smaller units.
[0009] Other features and advantages of the invention will become
apparent from the following detailed description, taken in
conjunction with the accompanying drawings which illustrate, by way
of example, various features of embodiments of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] FIG. 1 shows a glucose binding competitive affinity assay
based on Forster Resonance Energy Transfer (FRET) in accordance
with an embodiment of the invention;
[0011] FIG. 2 shows instrumentation for measuring the lifetime of
the excited state for a fluorophore-labeled assay in accordance
with an embodiment of the invention;
[0012] FIG. 3 shows a glucose binding competitive affinity assay,
including a reference fluorophore, for intensity interrogation in
accordance with an embodiment of the invention;
[0013] FIG. 4A shows instrumentation for interrogating a
fluorophore-labeled assay with an internal reference used for an
intensity interrogation in accordance with an embodiment of the
invention;
[0014] FIG. 4B shows instrumentation for interrogating a
fluorophore-labeled assay with an internal reference used for an
intensity interrogation in accordance with another embodiment of
the invention;
[0015] FIG. 5A is a perspective view of an optical fiber sensor
according to an embodiment of the invention;
[0016] FIG. 5B is a side view of the optical fiber sensor shown in
FIG. 5A;
[0017] FIG. 6A is a perspective view of an optical fiber sensor
with a compound parabolic concentrator (CPC)-shaped fiber tip a
according to an embodiment of the invention;
[0018] FIG. 6B is a side view of the optical fiber sensor shown in
FIG. 6A;
[0019] FIG. 7A is a perspective view of an optical fiber sensor
with a compound parabolic concentrator (CPC)-shaped fiber tip, a
reflector, and support structure in accordance with an embodiment
of the invention;
[0020] FIG. 7B is a side view of the optical fiber sensor shown in
FIG. 7A;
[0021] FIG. 7C is a side view of an optical fiber with side cut
cavities in accordance with an embodiment of the invention;
[0022] FIG. 7D is a perspective view of the optical fiber sensor
shown in FIG. 7C;
[0023] FIG. 8 shows the fluorescence distribution inside the assay
compartment of an optical fiber sensor with a straight-cut optical
fiber (height=diameter of assay compartment);
[0024] FIG. 9 shows the fluorescence distribution inside the assay
compartment of an optical fiber sensor with a CPC-shaped optical
fiber (height=diameter of assay compartment);
[0025] FIG. 10A shows the fluorescence distribution inside the
assay compartment of an optical fiber sensor with a CPC-shaped
optical fiber and reflector (height=diameter of assay
compartment);
[0026] FIG. 10B shows the fluorescence distribution inside the
assay compartments of an optical fiber sensor with side cut
cavities (height=diameter of fiber);
[0027] FIG. 11 shows a stacked planar integrated optical system
(SPIOS) for lifetime interrogation of a fluorophore-labeled assay
in accordance with an embodiment of the invention;
[0028] FIG. 12 shows the spectrum of light source, excitation
filter, and fluorophore for a lifetime system in accordance with an
embodiment of the invention;
[0029] FIG. 13 shows a stacked planar integrated optical system
(SPIOS) for intensity interrogation of a fluorophore-labeled assay
in accordance with an embodiment of the invention;
[0030] FIG. 14 shows the spectrum of light source, excitation
filter, emission filter, assay fluorophore, and reference
fluorophore for a intensity system in accordance with an embodiment
of the invention;
[0031] FIG. 15 shows a stacked planar integrated optical system
(SPIOS) for intensity interrogation of a fluorophore-labeled assay
in accordance with another embodiment of the invention;
[0032] FIGS. 16A and 16B show examples of a CPC SPIOS-fiber
interface in accordance with embodiments of the invention; and
[0033] FIG. 17 shows illustrative layers of a wafer-scale stacked
planar integrated optical system (SPIOS) in accordance with an
embodiment of the invention.
DETAILED DESCRIPTION
[0034] In the following description, reference is made to the
accompanying drawings which form a part hereof and which illustrate
several embodiments of the present invention. It is understood that
other embodiments may be utilized and structural and operational
changes may be made without departing from the scope of the present
invention.
[0035] As shown in the drawings for purposes of illustration,
embodiments of the invention are directed to optical sensors that
may be interrogated by optical, or optoelectronic, systems. Optical
sensors may be introduced and/or lodged transdermally, or may be
implanted in and/or through subcutaneous, dermal, sub-dermal,
inter-peritoneal, or peritoneal tissue. In the discussion herein,
preferred embodiments of the devices, systems, and methods of the
invention are described with reference to glucose as the analyte
whose level/concentration in the blood and/or bodily fluids of the
user is to be determined. However, this is by way of illustration
and not limitation, as the principles, devices, systems, and
methods of the present invention may be used for sensing and/or
determining the level of a variety of other physiological
parameters, agents, characteristics, and/or compositions.
[0036] As will be described in more detail below, an optical
glucose sensor having an assay compartment may be formed, e.g., by
including a glucose permeable membrane containing the assay at the
distal end of an optical fiber. The optical fiber may then be
inserted transdermally into the user's body, thereby situating the
assay compartment in the user's tissue, while leaving at least a
part of the optical fiber outside the body such that it can be
accessed by an interrogating system. Alternatively, the optical
sensor may be implantable, e.g., as part of an implantable glucose
monitor including an interrogating optoelectronic system and a
power source. The assay compartment may be formed between a glucose
permeable membrane and an optical interface to the optoelectronic
system. The optical sensor may preferably be biodegradable.
[0037] As shown in FIG. 1, an optical glucose sensor may be based
on a competitive glucose binding affinity assay. The assay may
include a glucose receptor and a glucose analog (ligand) contained
in a compartment where at least a part of the compartment is
capable of exchanging small molecules, such as glucose, salts, etc.
with the surrounding medium, while retaining macromolecules, such
as the assay components.
[0038] Several molecules may serve as the glucose receptor in the
glucose assay. Examples include, but are not limited to,
Concanavalin A, periplasmic glucose/galactose-binding receptor,
antibodies raised against glucose-like molecules, Boronic Acids,
and Mannan Binding Lectin (MBL). Mannan Binding Lectin is human
protein, which is a part of the innate immune system. Thus, the
assay may include MBL as the glucose receptor and dextran as the
glucose analog.
[0039] The binding between MBL and glucose-like molecules (e.g.,
dextran) is reversible. When no glucose is present, MBL and dextran
will predominantly be bound together. When glucose is added to the
assay, it will compete off a part of the dextran population, such
that the assay enters a new equilibrium state. The equilibrium
state at all times corresponds to the glucose concentration. In
order to determine this equilibrium state, MBL is labeled with a
fluorophore (e.g., Alexa Fluor 594, or AF594), and the dextran is
labeled with a dye (e.g., hexamethoxy crystalviolet-1 (HMCV1)--a
proprietary crystal violet derivative, Medtronic, Inc.). The donor
fluorophore and the acceptor dye together form a Forster Resonance
Energy Transfer (FRET) pair--i.e., the emission spectrum of the
fluorophore and the absorption spectrum of the dye overlap.
[0040] The occurrence of FRET affects the lifetime of the excited
state and the intensity of the emitted fluorescence and can only
occur when the fluorophore and the corresponding dye are in close
proximity (i.e., in the range of about 50.ANG.). Thus, the FRET
mechanism permits interrogation of the equilibrium state optically
by illuminating the assay and measuring either the lifetime of the
excited state, and/or the intensity of the emitted fluorescence
from the donor fluorophore. It is noted that the donor fluorophore
and the acceptor dye are preferably water soluble, as they are to
function in an aqueous environment.
[0041] FIG. 2 shows instrumentation used for frequency domain
lifetime interrogation of the above-described assay based on a
modified epifluorescence microscope. The instrumentation, or
optical interrogation system, is optically coupled to (or aligned
with) a sensor 100 carrying the assay. The assay is excited with a
periodic signal (e.g., sinusoidal, squarewave, dirac pulse,
approximative dirac, sawtooth, etc.), and the modulation frequency
is governed by the lifetime of the excited state (.tau.) for the
fluorophore. The optimum modulation frequency may be approximated
by:
f.sub.opt=1/(2*.pi.*.tau.) Eq. (1)
Thus, for a lifetime of 3 ns, e.g., the optimum modulation
frequency (f.sub.opt) is in the range of about 50 MHz to about 60
MHz.
[0042] With reference to FIG. 2, an oscillator 105 in combination
with a driver circuit 110 modulates a LED 120 with a wavelength
range capable of exciting the fluorophore. The LED 120 output is
filtered using a multilayer dielectrical filter 130 to select a
distinct wavelength region. The filtered LED output is reflected by
a dichroic beam splitter 140 and focused onto the sensor 100 (which
contains the assay) by a lens 150. The assay emits fluorescence
with the same frequency as the excitation (modulated LED output)
and phase shifted as a result of the lifetime of the excited state
for the fluorophore.
[0043] The emitted fluorescence 103 and the reflected excitation
light 123 are picked up and collimated by the lens 150. The
dichroic beam splitter 140 transmits the fluorescence 103. However,
it reflects the majority of the back-reflected excitation light
123. An emission filter 160 with a distinct wavelength region red
shifted with respect to, and not overlapping, the pass band of the
excitation filter blocks the remaining part of the excitation light
123 and transmits the fluorescence 103. Thus, in effect, only the
fluorescence carrying the modulated and phase shifted fluorescence
is focused onto a photodetector 180 using a lens 170. The phase lag
between the detected fluorescence and the excitation light
correlates with the glucose concentration in the assay.
[0044] In addition to the lifetime of the excited state, the
intensity of the emitted fluorescence also correlates to the
glucose concentration. In contrast to a lifetime measurement, the
measured intensity of the emitted fluorescence is affected by the
intensity of the light source and the coupling between the assay
and the optical system. Therefore, the intensity measurement
requires an internal reference fluorophore to be incorporated into
the assay, as shown in FIG. 3.
[0045] The reference fluorophore must differ from the assay
fluorophore in a way that the emitted fluorescence from the assay
and that from the reference may be separated from one another,
e.g., by having different absorption spectra or emission spectra.
The reference fluorophore may be, e.g., Alexa Fluor 700 (AF700)
labeled onto Human Serum Albumin (HSA) or another macro molecule,
which largely does not bind to the glucose receptor. See FIG. 3.
Alexa Fluor 700 may be excited simultaneously with the Alexa Fluor
594 as their absorption spectra spectrally overlap. The emission
spectrum from Alexa Fluor 700 is slightly red shifted with respect
to Alexa Fluor 594, which makes it possible to detect their
respective fluorescence emissions in separate wavelength regions.
As they are excited simultaneously by the same light source, any
changes in the intensity of the light source will scale
fluorescence from AF594 and AF700 equally. As such, any effect
originating from changes in the intensity of the light source may
be cancelled out.
[0046] The excitation, as well as the detection, of the emitted
fluorescence for the assay and the reference follow the same
optical path from the optical system to the assay. As such, the
detected signal from the reference serves as a measure for the
optical coupling between the optical interrogating system and the
assay. Any effect originating from changes in the optical coupling
such as alignment may be cancelled out.
[0047] FIG. 4A shows one embodiment of instrumentation used for
fluorescence interrogation of the above-described assay based on
another modification of an epifluorescence microscope. A driver
circuit 310 modulates a LED 320 at a low frequency--solely with the
purpose of eliminating the 1/f noise and canceling out ambient
light--with a wavelength range capable of simultaneously exciting
the assay and reference fluorophores. The LED output is filtered
using a multilayer dielectrical filter 330 to select a distinct
wavelength region. The filtered LED output is reflected by a first
dichroic beam splitter 340 and focused onto the sensor 300, which
includes the assay and the reference, by a lens 350.
[0048] The assay and the reference emit fluorescence. The emitted
fluorescence 301 and the reflected excitation light 323 are picked
up and collimated by the lens 350. The first dichroic beam splitter
340 transmits the fluorescence 301. However, it reflects the
majority of the back reflected excitation light 323. A second beam
splitter 344 reflects the reference fluorescence at a 90.degree.
angle 307, but it transmits the assay fluorescence 309. An assay
emission filter 360 with a distinct wavelength region red shifted
with respect to, and not overlapping, the pass band of the
excitation filter and matching the desired part of the assay
fluorescence spectrum then blocks the remaining part of the
excitation light and transmits the assay fluorescence.
[0049] Similarly, a reference emission filter 364 with a distinct
wavelength region red shifted with respect to, and not overlapping,
the pass band of the excitation filter and matching the desired
part of the reference fluorescence blocks the remaining part of the
excitation light and transmits the reference fluorescence 307.
Thus, in effect, only the fluorescence from the assay and the
fluorescence from the reference are focused onto their respective
photo detectors 380, 384 using respective lenses 370, 374. The
ratio between the detected assay fluorescence and the detected
reference fluorescence correlates with the glucose concentration in
the assay. As mentioned previously, any changes in light-source
intensity or optical coupling will be cancelled out as they scale
the assay and reference fluorescence equally.
[0050] FIG. 4B shows another embodiment of the instrumentation used
for fluorescence interrogation. Here, as in FIG. 4A, driver circuit
310 modulates a LED 320 at a low frequency--solely with the purpose
of eliminating the 1/f noise and canceling out ambient light--with
a wavelength range capable of simultaneously exciting the assay and
reference fluorophores. The LED output is filtered using a
multilayer dielectrical filter 330 to select a distinct wavelength
region. The filtered LED output is reflected by a first dichroic
beam splitter 340 and focused onto the sensor 400, which includes
the assay and the reference, by a lens 350. The sensor 400 is a
fiber optical sensor, as described more fully hereinbelow.
[0051] As described in connection with FIG. 4A, the assay and the
reference emit fluorescence. The emitted fluorescence 301 and the
reflected excitation light 323 are picked up and collimated by the
lens 350. The first dichroic beam splitter 340 transmits the
fluorescence 301. However, it reflects the majority of the back
reflected excitation light 323. A second beam splitter 344 reflects
the assay fluorescence at a 90.degree. angle 309, but it transmits
the reference fluorescence 307. A reference emission filter 364
with a distinct wavelength region red shifted with respect to, and
not overlapping, the pass band of the excitation filter and
matching the desired part of the reference fluorescence spectrum
then blocks the remaining part of the excitation light and
transmits the reference fluorescence.
[0052] Similarly, an assay emission filter 360 with a distinct
wavelength region red shifted with respect to, and not overlapping,
the pass band of the excitation filter and matching the desired
part of the assay fluorescence blocks the remaining part of the
excitation light and transmits the assay fluorescence 309. Thus, in
effect, only the fluorescence from the assay and the fluorescence
from the reference are focused onto their respective photo
detectors 380, 384 using respective lenses 370, 374. The ratio
between the detected assay fluorescence and the detected reference
fluorescence correlates with the glucose concentration in the
assay. Again, as mentioned previously, any changes in light-source
intensity or optical coupling will be cancelled out as they scale
the assay and reference fluorescence equally.
[0053] FIGS. 5A and 5B show an embodiment of the invention, wherein
a fiber optical sensor 400 is made by placing the assay in a
compartment 420 that is distal to the distal end 412 of an optical
fiber 410. In this embodiment, a test tube-shaped glucose permeable
membrane 430 containing the assay is slid over the end of the
optical fiber 410 and sealed (e.g., heat sealed). The distal end
412 of the fiber 410 is straight cut and polished, and is in direct
contact with the assay. In embodiments of the invention, the
glucose permeable membrane 430 may be made of a biocompatible,
biodegradable polymer such as, e.g., PolyActive.TM. (Integra
Orthobiologics, Irvine, Calif.), Poly-lactide-glycolic-acid,
poly-caaprolactone and non-biodegradable polymers exhibiting
molecular weight cut-off properties like Cellulose (Spectrum
Laboratories, Rancho Dominguez, Calif.) or Polysulfone (Spectrum
Laboratories, Rancho Dominguez, Calif.).
[0054] With the above configuration, the assay may now be excited
through the optical fiber 410, and the resulting fluorescence
collected by the optical fiber. As the fluorescence from the assay
radiates isotropically, the amount of the emitted fluorescence,
which can be picked up by the optical fiber 410, is set by the
numerical aperture of the fiber.
[0055] The numerical aperture (NA) of the optical fiber is a
function of the refractive index of the fiber core (n.sub.1) and
the refractive index of the cladding (n.sub.2):
NA= {square root over (n.sub.1-n.sub.2.sup.2)} Eq. (2)
[0056] Generally, light entering the optical fiber at an angle less
than a critical angle will be transmitted through the optical fiber
due to total internal reflection in the core/cladding boundary,
whereas light entering at an angle larger than the critical angle
will simply exit the fiber through the cladding. Commercially
available optical fibers have a high refractive index core and a
low refractive index cladding. Typical refractive indices for the
core and the cladding for a plastic optical fiber are about 1.49
and about 1.40, respectively, which, based on Eq. (2), results in a
numerical aperture of about 0.51. Per Eq. (3) below, this
corresponds to a critical angle (.theta.) of about 30.6.degree. ,
or a solid angle of about 0.88sr:
NA=n sin.theta..sub.max Eq. (3)
[0057] For the ideal case, this translates into about a 7% pickup
of the total emitted fluorescence--where isotropic radiation is
4.pi.sr, and 0.88sr/4.pi.sr.apprxeq.7%. The maximum fluorescence
pickup is thus set by the optical fiber.
[0058] Furthermore, some of the excitation light will spill out of
the assay compartment through the glucose permeable membrane as the
excitation light is coupled into the assay at angles corresponding
to the fiber numerical aperture.
[0059] The Optical Invariant theorem states that the product of the
source area (A) multiplied by the solid angle (.OMEGA.) is
constant:
A.sub.1.OMEGA..sub.1=A.sub.2.OMEGA..sub.2 Eq. (4)
[0060] In Eq. (4), A1 is equal to the cross sectional area of the
optical fiber, .OMEGA.1 equals the solid angle corresponding to the
numerical aperture of the optical fiber, and A2 and A.OMEGA.2 are
set according to a trade-off between the maximum fiber tip length,
saturation intensity for the assay, and manufacturable tip
geometry.
[0061] Applied to the fiber sensor 400, this means that, with the
right optical component and fiber tip design, the excitation light
transmitted through the optical fiber 410 may be focused down to a
smaller area moving the excitation away from the glucose permeable
membrane 430 and thereby reduce the spillage of excitation light
out of the sensor. In addition, the numerical aperture of the fiber
tip may be increased resulting in an increased fluorescence pickup
from the assay.
[0062] The Compound Parabolic Concentrator (CPC) is a non-imaging
component, which has an entrance aperture, a parabolic shaped
reflective surface, and an exit aperture. The CPC may be formed as
an air filled void or an optical material with a parabolic mirror
surface, or it may be formed by an optical material with a
refractive index which is higher than that of the surrounding
material. The parabolic shaped part of the CPC is formed as a
parabola which ensures total internal reflection due to the
high-low refractive index transition.
[0063] The radial coordinate of points on the CPC as a function of
the z coordinate along the axis is given by the positive real root
of the following quadratic equation:
C.sup.2r.sup.2+2(CSz+aP.sup.2)r+(z.sup.2S.sup.2-2aCQz-a.sup.2PT)=0
Eq. (5)
Where C=cos.theta., S=sin.theta., P=1+S, Q=1+P, and T=1+Q
[0064] Shaping the tip of the optical fiber 410 as a CPC with the
right dimensions will lead to the desired properties. As an
example, a CPC shape applied to the tip of a 250 .mu.m optical
fiber reduces tip diameter to 125 .mu.m. Area is thus reduced four
times, leading to a theoretical four-fold increase in numerical
aperture, which corresponds to a four-fold increase in fluorescence
pickup.
[0065] An embodiment of the present invention employing a CPC tip
geometry is shown in FIGS. 6A and 6B. As shown, the CPC shaped
fiber tip 414 is in direct contact with the assay, which has a
refractive index similar to water (1.33). As this is significantly
lower than the refractive index of the optical fiber cladding, a
CPC designed to have a cladding on the parabolic part will work
with, as well as without, cladding.
[0066] It is noted that the theoretical four-fold increase in
fluorescence pickup is based on the assumption that the
fluorophore(s) of the assay are excited at the tip-assay
transition, i.e., the CPC-assay optical interface 415. However,
since the fluorescence occurs in a volume in front of the fiber tip
414, the increase in fluorescence pickup may be significantly less
than the four-fold increase predicted by crude theoretical
calculations.
[0067] Thus, even though, theoretically, the numerical aperture of
the CPC fiber tip 414 will be increased dramatically compared to
the conventional straight cut fiber tip 412, in operation, the CPC
design generally cannot pick up more than about 50% of the emitted
fluorescence. Nevertheless, in an embodiment of the invention, the
fluorescence pick-up percentage may be increased by placing a
concave mirror 417 in front of the fiber tip 414 to reflect
fluorescence emitted in a direction that is opposite to the fiber
tip and focus it into the fiber.
[0068] As shown in FIGS. 7A and 7B, there must still be a gap
between the fiber tip 414 and the mirror 417 to leave room for the
assay, i.e., to constitute the assay compartment 420. The mirror
417 obviously needs to be kept in place in front of the fiber tip
414. This may be accomplished, e.g., by the support structure 419,
which allows glucose to diffuse into the space in front of the
fiber tip 414. At the same time, the support structure 419 serves
the purpose of supporting the glucose permeable membrane 430.
[0069] The straight cut fiber tip 412, the CPC shaped fiber tip
414, and the CPC shaped fiber tip in combination with a reflector
414, 417 have been modeled using Zemax optical design software. For
each of the three designs, the model included the following: (1) an
excitation light source at the proximal (free) end of the fiber
sensor, coupling light into the optical fiber and exciting the
assay at the (distal, in-situ) tip of the optical fiber; (2) an
optical fiber with the selected fiber tip geometry; (3) an
assay-filled compartment including assay absorption and assay
fluorescence processes; and (4) a detector with a fluorescence
filter only selecting the fluorescence picked up and transmitted
back through the optical fiber.
[0070] For all three designs, the ratio between the excitation and
the detected fluorescence was calculated. As can be seen from the
results shown in Table 1 below, the Zemax simulations shows
significantly lower fluorescence pickup from the CPC designs than
the Optical Invariant theorem predicts. As stated, this is due to
the fact that the excitation and resulting fluorescence emission
occur in a volume in front of the fiber tip rather than directly at
the fiber tip/assay boundary.
TABLE-US-00001 TABLE 1 Without Reflector With Reflector Straight
Cut 100% N/A CPC 166% 277%
[0071] In embodiments of the invention employing the CPC tip
geometry described above, the assay, including the reference dye,
can be carried or contained by a hydrogel in order to ease
production and stabilize the assay. Specifically, the glucose assay
is first dissolved in a hydrogel. Next, the CPC-shaped fiber tip
may be dipped into the hydrogel containing the assay, and a droplet
may be left in front of the CPC-shaped fiber tip. Finally, the
hydrogel may be cross-linked so as to provide a glucose sensor.
Suitable cross-linking hydrogels may include, e.g., Poly acryl
based hydrogels (such as poly-Hydroxy-Ethyl-Methacrylate (pHEMA),
PMMA-pHEMA co-polymers, etc.), Polyurethanes, Polyesters,
Polyethers, etc.
[0072] Where the hydrogel containing the assay is not cross-linked,
the entire embodiment could be coated by any of the polymers
suitable for glucose sensors, i.e., polymers that allow glucose
diffusion through the polymer. Suitable non-crosslinking hydrogels
may include, e.g., poly-vinyl alcohol (PVA), Poly-ethylene glycol
(PEG), poly-propylene glycol (PPG), poly-Hydroxy-Ethyl-Methacrylate
(pHEMA), etc., and co-polymers thereof.
[0073] FIGS. 7C and 7D show another embodiment of the invention,
wherein a fiber optical sensor 1400 is made by placing the assay in
the compartments 1420a, 1420b proximate the distal end 1412 of an
optical fiber 1410. The compartments are formed as side cuts in the
fiber cutting through the fiber cladding as well as core. In this
embodiment, a tube-shaped glucose permeable membrane 1430 is slid
over the side cuts and sealed to the cladding of the fiber leaving
glucose permeability in the regions of the assay compartments.
[0074] Two cuts are made on opposite sides of the fiber and
displaced from each other sufficiently to maintain the structural
strength of the fiber. For the first side cut cavity 1420a, the
surface 1421 a parallel to the fiber axis is preferably optical
quality. As the refractive index of the assay is significantly
lower than the fiber core, this will provide total internal
reflection for excitation light 1423 traveling from the proximal
end of the fiber sensor to the second assay compartment 1420b and
furthermore provide total internal reflection for fluorescence
emitted from the second assay compartment 1420b and back to the
proximal end of the fiber. The above-described configuration
provides structural strength to the fiber sensor, which is
advantageous, especially for fiber sensors where a soft glucose
permeable membrane in itself does not provide sufficient structural
strength to ensure the stability of the assay compartment.
[0075] It is noted that, in embodiments of the invention, fewer or
more assay compartments may be included. For example, with
reference to FIGS. 7C and 7D, the second assay compartment 1420b
may be omitted when, e.g., a single assay compartment provides
sufficient fluorescence. Also, in additional embodiments, as an
alternative to the combination of a liquid assay and a glucose
permeable membrane, a hydrogel with embedded assay as described
above may be cast into the one or more side cavities to form the
assay compartment(s).
[0076] FIGS. 8-10A show the fluorescence intensity distribution
inside the assay compartment 420 for each of the aforementioned
designs. Clearly, while, with the straight cut fiber, the
fluorescence emission zone extends out to the membrane, the CPC
concentrates the excitation and the fluorescence emission in the
center of the assay compartment. FIG. 10B shows the fluorescence
intensity distribution inside the assay compartments for the side
cut cavities shown in FIGS. 7C and 7D.
[0077] It is noted that the CPC configuration applied to the fiber
sensor in the above description is illustrative, and other
geometries, e.g., a CPC with a rectangular cross sectional area, as
well as other imaging or non-imaging geometries that also change
the numerical aperture of the fiber tip may also be applied.
[0078] Embodiments of the invention are also directed to improved
optical systems for interrogating fluorophore-labeled assays
contained within optical sensors and, in particular, fiber optical
sensors of the types described above.
[0079] As noted previously, in typical interrogation systems (e.g.,
epifluorescence microscopy), fluorescence applications commonly
operate with fairly intense excitation of the fluorophore in the
absorption band of the fluorophore and detection of the weak
fluorescence emitted by the fluorophore. In such applications, the
dichroic beam splitter serves as a crude filtering of the light,
whereas the heavy filtering occurs in the excitation filter and the
emission filter, which are used in transmission mode and depend on
10.sup.6 times attenuation of wavelengths outside the pass band in
a single pass. Such filters are normally based on dielectric
multilayer filters consisting of a substrate with optical coatings
on both sides consisting of up to 100 layers with alternating
refractive indices. In the pass band, the filters have up to 99%
transmittance, which is most probably caused by reflection losses
in the air-coating transition on both sides of the filter in spite
of the fact that filter stacks on both sides of the substrate
generally include an anti-reflective coating.
[0080] In accordance with embodiments of the present invention, an
optical system may be used for interrogating a fluorophore-labeled
assay either with, or without, an internal reference. The inventive
optical system is based on a filter substrate with one or multiple
optical coatings separated in location on the surfaces of the
substrate. The coatings may be dielectrical multilayer coatings
forming short wave pass, longwave pass, band pass filters, and
anti-reflection coatings. Furthermore, coatings may be metallic
reflective coatings.
[0081] More specifically, embodiments of the present invention
utilize the fact that a dielectrical multilayer filter reflects
what is not transmitted. Therefore, a filter in accordance with
embodiments of the present invention may include a first coating on
the filter substrate which transmits a certain wavelength range of
the excitation, but reflects wavelengths outside the pass band.
This enables the emitted fluorescence to be reflected once or
multiple times by the coating, while allowing reflected excitation
light to be transmitted out of the optical system. The filtered
fluorescence then exits the filter substrate when it reaches a part
of the substrate that is not coated with the first coating, and is
picked up by one or more detectors.
[0082] Subsequent filtering may also be achieved by applying a
second coating which transmits a desired wavelength range
originating from a first fluorophore, but reflects wavelengths
outside the transmission band--in particular, remains of the
excitation light. In this way, the desired wavelength range will be
transmitted out of the substrate where it may be picked up by one
or more suitable detectors. In addition, anti-reflective coatings
may be applied to areas of the substrate where light is coupled
into, or out of, the substrate to reduce reflection losses.
[0083] In an intensity interrogation configuration, where both a
first fluorophore and a second fluorophore are used, the
above-mentioned second coating may transmit the desired wavelength
range associated with the first fluorophore and reflect wavelengths
outside the transmission band--in particular remains of the
excitation light and fluorescence related to the second
fluorophore. The filtered fluorescence originating from the second
fluorophore then exits the filter substrate when it reaches a part
of the substrate that is not coated with the first or second
coating, such that it can be picked up by a suitable detector(s).
Subsequent filtering of fluorescence originating from the second
fluorophore may be achieved by applying a second coating which
transmits the desired wavelength range associated with the second
fluorophore and reflects wavelengths outside the transmission
band--in particular, remains of the excitation light. In this way,
the desired wavelength range may be transmitted out of the
substrate where it may be picked up by a suitable detector(s). In
other embodiments, the system may be expanded to include multiple
light sources and/or multiple wavelength ranges to be detected.
[0084] Implementation of the above-described filter configurations
requires imaging optical elements such as lenses, mirrors or
diffractive optical elements to focus, pick up, and collimate
light. Furthermore, apertures and light traps may be required to
control light path and to absorb undesired wavelengths that are
transmitted out of the filter substrate, such as, e.g., the
excitation light to be blocked. In this embodiment, the
above-mentioned optical elements and light traps are shown with an
air gap between the elements and the (glass) substrate. However,
the optical elements and light traps may also be formed in an
optically transparent material with mirror coating and absorbing
coating on the surface facing away from the filter substrate. Such
a configuration is generally more advantageous, as the optical
elements and light traps are better index matched to the filter
substrate/filter coating than is the case when an air gap is
present.
[0085] The above-mentioned elements may be individually aligned and
fitted onto both sides of the coated filter substrate as discrete
units. Similarly, one or more light sources and one or more
detectors may be aligned and fitted onto the filter substrate as
packaged units, as raw dies laminated directly onto the coated
filter substrate, or mounted as raw dies onto a printed circuit
board and mounted as a unit onto the coated filter substrate.
[0086] Suitable light sources may include, e.g., light emitting
diodes (LEDs) and laser diodes, and suitable detectors may include,
e.g., photodiodes, avalanche photodiodes, silicon photomultipliers,
photomultipliers, and phototransistors. In addition, the assembled
optical system may be coated or placed in an enclosure to block out
ambient light.
[0087] A stacked planar integrated optical system (SPIOS) for
interrogating a single fluorophore in accordance with an embodiment
of the invention is shown in FIG. 11. With reference to FIGS. 11
and 12, a LED 510 emits light into a filter substrate 500 with a
wavelength range overlapping the absorption spectrum of the
fluorophore to be interrogated. The LED output is limited to a
certain wavelength range by an excitation filter 520 before
entering the filter substrate 500. The filtered excitation light
exits the filter substrate 500 through an identical excitation
filter on the opposite side of the filter substrate and is
collimated by a first mirror 530. The collimated excitation light
passes through the filter substrate to reach a second mirror 540
and is thereby focused onto a sensor 590 through an optical window
550. It is noted that, in this embodiment, the sensor 590 is a
fiber optical sensor--including an assay in an assay compartment
595--of the types described above in connection with FIGS.
5-10.
[0088] Traveling through the sensor 590, the excitation light 591
reaches the assay compartment 595, where it excites the fluorophore
in the assay such that the fluorophore emits fluorescence 593.
Furthermore, excitation light is reflected and back scattered from
the optical window and sensor. The fluorescence 593 and
reflected/back scattered excitation light are picked up and
collimated by the second mirror 540 and enter the filter substrate
500 through an uncoated or anti-reflection coated area 503 on the
filter substrate. The emitted fluorescence is reflected between the
two coated surfaces (i.e., the excitation filters 520) while the
excitation light is transmitted through the coatings 520 and
absorbed by the light traps 560. The filtered fluorescence exits
the filter substrate 500 where the coated area ends 507 and is
focused by a third mirror 570 onto a detector 580 through an
uncoated, or anti-reflection coated, region of the filter
substrate.
[0089] In an alternative embodiment, a stacked planar integrated
optical system (SPIOS) for interrogating a sensor with an assay
fluorophore and a reference fluorophore is shown in FIG. 13. With
reference to FIGS. 13 and 14, a LED 510 emits light into a filter
substrate 500 with a wavelength range overlapping the absorption
spectrum of the two fluorophores to be interrogated. The LED output
is limited to a certain wavelength range by an excitation filter
520 before entering the filter substrate 500. The filtered
excitation light exits the filter substrate 500 through an
identical excitation filter on the opposite side of the filter
substrate and is collimated by a first mirror 530. The collimated
excitation light passes through the filter substrate to reach a
second mirror 540 and is thereby focused onto a sensor 590 through
an optical window 550. It is noted that, in this embodiment, the
sensor 590 is a fiber optical sensor--including an assay in an
assay compartment 595--of the types described above in connection
with FIGS. 5-10.
[0090] Traveling through the sensor 590, the excitation light 591
reaches the assay compartment 595, where it excites the fluorophore
in the assay such that the fluorophore emits fluorescence 593.
Furthermore, excitation light is reflected and back scattered from
the optical window and sensor. The fluorescence 593 and
reflected/back scattered excitation light are picked up and
collimated by the second mirror 540 to enter the filter substrate
500 through an uncoated or anti-reflection coated area 503 on the
filter substrate. The emitted fluorescence is reflected between the
two excitation filter coatings 520 while the excitation light is
transmitted through the coatings and absorbed by the light traps
560.
[0091] The filtered fluorescence exits the filter substrate 500
where the excitation filter coatings end 507, and an emission
filter 525 transmits a wavelength range relating to a first
fluorophore (the assay fluorophore). The filtered fluorescence from
the first fluorophore is focused by a third mirror 570 onto a first
detector 580, while the fluorescence associated with the second
fluorophore (the reference fluorophore) is reflected between the
emission filter 525 on the two sides of the filter substrate 500.
The filtered fluorescence from the second fluorophore exits the
filter substrate 500 where the coated area ends 509 or is replaced
with an anti-reflection coating, and is focused by a fourth mirror
575 onto a second detector 585 through an uncoated, or
anti-reflection coated, region of the filter substrate.
[0092] As noted previously, the optical elements, apertures, and
light traps of the SPIOS for interrogating a sensor with an assay
fluorophore and a reference fluorophore may be disposed with an air
gap between the elements and the (glass) substrate. See FIG. 13.
However, the optical elements and light traps may also be formed in
an optically transparent material with mirror coating and absorbing
coating on the surface facing away from the filter substrate. The
latter configuration is generally more advantageous, as the optical
elements and light traps are better index matched to the filter
substrate/filter coating than is the case when an air gap is
present. An example of such a configuration is shown in FIG. 15,
where connecting material 1501 is provided between the optical
components.
[0093] It is noted that the light emitting area of a LED chip has
an area, which is comparable to a 500 .mu.m multimode fiber.
Furthermore, the LED chip emits in a large angular space. For a
SPIOS equipped with a LED interrogating a fiber sensor, the
fluorescence output gradually becomes limited by the ability to
focus light from the LED onto the fiber when the fiber diameter is
reduced. Also the positioning of the proximal end of the fiber
relative to the optical system becomes more critical.
[0094] As shown in FIGS. 16A and 16B, the ability to couple light
from the LED into the fiber sensor may be enhanced by placing a
compound parabolic concentrator (CPC) 1505 in the interface between
the SPIOS and the fiber sensor 1590 if the numerical aperture of
the SPIOS in relation to the fiber is smaller than the numerical
aperture of the fiber itself. The CPC may then be used to match the
numerical aperture of the fiber and thereby reduce the spot size of
the focused excitation light originating from the LED. This, in
turn, enables more light to be coupled from the LED into the
fiber.
[0095] The CPC may be an integral part of the SPIOS, as shown,
e.g., in FIG. 16A. However, the CPC may also be formed on the
proximal end of the fiber, whereby the area to be targeted by the
SPIOS is increased, thus reducing the requirements on the
positioning of the fiber (with CPC at the proximal end) relative to
the SPIOS.
[0096] In yet another embodiment shown in FIG. 17, the inventive
optical interrogating system may be designed to be manufactured as
a wafer-scale stacked planar integrated optical system, or
wafer-scale SPIOS (also referred to as a "Wafer Scale Optical
System" or a "Wafer Level Optical System"). As shown in FIG. 17,
the SPIOS includes various layers that are stacked and aligned. In
the wafer layer 610, one or more light sources (e.g., LEDs and
photodiodes) and detectors may be laid out on a wafer.
Alternatively, they may be naked chips (e.g., sold by Avago
Technologies or Hamamatsu), which are individually aligned and
laminated onto the SPIOS units.
[0097] One or more optical layers 620 may include mirrors and
absorbers laid out on a wafer-sized injection molded disk. Mold
inserts defining optical surfaces are made by a diamond
turning/milling company (e.g., Kaleido Technology in Denmark). Gold
or protected silver is applied to mirror surfaces, e.g., by
sputtering, while any absorbers are masked off during the
process.
[0098] The optical filter layer 630 includes a wafer-sized glass
substrate with optional coatings. Specifically, multilayer optical
coatings may be applied on both sides of the glass substrate using
ion-assisted sputtering to form durable coatings. The technique is
similar to that used in manufacturing fluorescence filters by,
e.g., Semrock in the United States and Delta in Denmark.
[0099] As shown in FIG. 17, in one embodiment, a wafer layer 610
may be followed by an optical layer 620, an optical filter layer
630, and another optical layer 620. The entire stack is then
thoroughly aligned and laminated, e.g., by gluing, and the
connections are bonded onto the chips. The stack is then diced 640
using, e.g., a diamond saw to form multiple assembled SPIOS units
670.
[0100] The above-described system may be made small and is suitable
for large-scale production. The system may be used for
interrogating a sensor in a light scattering environment, such as a
sensor implanted into the skin, as well as a fiber sensor. Coating
or packaging may be used to block out ambient light.
[0101] While the description above refers to particular embodiments
of the present invention, it will be understood that many
modifications may be made without departing from the spirit
thereof. The accompanying claims are intended to cover such
modifications as would fall within the true scope and spirit of the
present invention.
[0102] The presently disclosed embodiments are therefore to be
considered in all respects as illustrative and not restrictive, the
scope of the invention being indicated by the appended claims, and
all changes which come within the meaning and range of equivalency
of the claims are therefore intended to be embraced therein.
* * * * *