U.S. patent application number 09/941807 was filed with the patent office on 2002-02-21 for optical sensor and method of operation.
This patent application is currently assigned to Bayer Corporation. Invention is credited to Mason, Richard W., Slovacek, Rudolf E., Sullivan, Kevin J..
Application Number | 20020020206 09/941807 |
Document ID | / |
Family ID | 21743840 |
Filed Date | 2002-02-21 |
United States Patent
Application |
20020020206 |
Kind Code |
A1 |
Mason, Richard W. ; et
al. |
February 21, 2002 |
Optical sensor and method of operation
Abstract
A multiple single use optical sensor includes a series of
continuous sensor stripes deposited on a substrate web. At least
one sample chamber is adapted to extend transversely across a
discrete portion of the series of sensor stripes to facilitate
analysis of a sample disposed therein. The sample chamber may be
moved, or additional sample chambers provided to enable subsequent
measurements of additional samples at unused discrete portions of
the sensor stripes. The continuous nature of the sensor stripes
provides consistency along the lengths thereof to enable
calibration data obtained from one discrete portion of the sensor
stripes to be utilized for testing an unknown sample an other
discrete portion of the sensor stripes. This advantageously
eliminates the need for any particular discrete portion of the
sensor stripes to be contacted by more than one sample, for
improved sensor performance.
Inventors: |
Mason, Richard W.; (Millis,
MA) ; Slovacek, Rudolf E.; (Norfolk, MA) ;
Sullivan, Kevin J.; (Medfield, MA) |
Correspondence
Address: |
WEINGARTEN, SCHURGIN, GAGNEBIN & LEBOVICI LLP
TEN POST OFFICE SQUARE
BOSTON
MA
02109
US
|
Assignee: |
Bayer Corporation
|
Family ID: |
21743840 |
Appl. No.: |
09/941807 |
Filed: |
August 29, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09941807 |
Aug 29, 2001 |
|
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09010096 |
Jan 21, 1998 |
|
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6306347 |
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Current U.S.
Class: |
73/1.02 ;
250/252.1; 436/8 |
Current CPC
Class: |
Y10T 436/10 20150115;
G01N 21/8483 20130101 |
Class at
Publication: |
73/1.02 ; 436/8;
250/252.1 |
International
Class: |
G01N 027/00; G01N
037/00 |
Claims
Having thus described the invention, what is claimed is:
1. An optical sensor adapted for sensing analyte content of a
plurality of samples, said optical sensor comprising: a substrate
web of predetermined length, said substrate web being substantially
gas impermeable and optically transparent in a predetermined
spectral range; a plurality of elongated sensor stripes extending
in parallel, spaced relation along the length of said web; each one
of said plurality of sensor stripes adapted for providing an
optically discernible response to presence of at least one analyte;
said optical sensor adapted for selective analyte-sensing contact
with the plurality of samples, wherein each one of the plurality of
samples are selectively superimposable with each one of said
plurality of elongated sensor stripes at one of a plurality of
discrete sample positions along the lengths thereof; said optically
discernible response being substantially identical at said
plurality of discrete sample positions.
2. The optical sensor as set forth in claim 1, wherein the
plurality of samples comprises at least one unknown sample and at
least one calibration sample, said optical sensor adapted for being
calibrated upon disposition of the calibration sample in said
analyte-sensing contact with said optical sensor at at least one of
said plurality of discrete sample positions distinct from that of
said at least one unknown sample.
3. The optical sensor as set forth in claim 1, wherein each one of
said plurality of sensor stripes exhibits said optically
discernible response in presence of incident light of a
predetermined spectral range.
4. The optical sensor as set forth in claim 1, further comprising a
multiple single use device, wherein each one of said discrete
sample positions along the lengths of said sensor stripes is
adapted for analyte-sensing contact with a single one of the
plurality of samples.
5. The optical sensor as set forth in claim 1, wherein the sample
is a fluid and said analyte-sensing contact comprises
surface-to-surface contact of the fluid with said sensor
stripes.
6. An optical sensor assembly adapted for sensing analyte content
of a plurality of samples, said optical sensor assembly comprising:
the optical sensor as set forth in claim 1; at least one sample
chamber superimposable with each of said plurality of elongated
sensor stripes at one of said plurality of discrete sample
positions along the lengths thereof; wherein said at least one
sample chamber is adapted for alternately maintaining individual
ones of the plurality of samples in said analyte-sensing
contact.
7. The optical sensor assembly as set forth in claim 6, wherein
said at least one sample chamber comprises: an elongated cavity
disposed within a chamber member, said elongated cavity being
defined by a substantially concave surface of said chamber member;
said elongated cavity including first and second apertures disposed
at opposite ends thereof to facilitate alternate entry and exit of
the individual ones of the plurality of samples to and from said
sample chamber; said chamber member adapted to extend across said
plurality of sensor stripes with said substantially concave surface
facing said web, wherein said optical sensor effectively closes
said substantially concave surface to define a longitudinal side
wall of said elongated cavity.
8. The optical sensor assembly as set forth in claim 7, further
comprising a plurality of said sample chambers.
9. The optical sensor assembly as set forth in claim 7, wherein
said at least one sample chamber is moveable for selective
superimposition with said plurality of discrete sample positions
along the lengths of said sensor stripes.
10. The optical sensor assembly as set forth in claim 9, wherein
said at least one sample chamber is adapted to extend orthogonally
to each of said plurality of elongated sensor stripes.
11. An optical sensor assembly adapted for sensing analyte content
of a plurality of samples, said optical sensor assembly comprising:
the optical sensor as set forth in claim 1; a plurality of sample
chambers disposed in parallel, spaced relation on said web, each
one of said plurality of sample chambers being sealably superposed
with said plurality of elongated sensor stripes at one of said
plurality of discrete sample positions along the lengths thereof;
wherein each of said plurality of sample chambers is adapted for
alternately maintaining individual ones of the plurality of samples
in said analyte-sensing contact.
12. The optical sensor as set forth in claim 11, wherein the
plurality of samples comprises at least one unknown sample and at
least one calibration sample, said optical sensor adapted for being
calibrated upon disposition of the calibration sample in one of
said sample chambers distinct from an other sample chamber adapted
to receive said at least one unknown sample.
13. The optical sensor assembly as set forth in claim 11, wherein
each of said plurality of sample chambers comprises: an elongated
cavity disposed within a chamber member, said elongated cavity
being defined by a substantially concave surface of said chamber
member; said elongated cavity including first and second apertures
disposed at opposite ends thereof to facilitate alternate entry and
exit of at least an individual one of the plurality of samples to
and from said sample chamber; said chamber member sealably
superposed with said substrate web and said plurality of sensor
stripes, wherein a discrete portion of said optical sensor
effectively closes said substantially concave surface to define a
longitudinal side wall of said elongated cavity.
14. The optical sensor assembly as set forth in claim 13, wherein
said chamber member further comprises: a chamber web sealably
superposed with said substrate web and said sensor stripes; a cover
web sealably superposed with said chamber web; said chamber web
having a plurality of slots extending in spaced parallel relation
across said sensor stripes; wherein each said slot and each portion
of said cover web superposed therewith define said concave
surface.
15. The optical sensor assembly as set forth in claim 14, wherein
said entry and exit apertures are disposed in said cover web.
16. The optical sensor assembly as set forth in claim 14, wherein
said entry and exit apertures are disposed in said substrate
web.
17. The optical sensor assembly as set forth in claim 14, wherein
at least one of said entry and exit apertures is disposed in said
substrate web and at least one of said entry and exit apertures is
disposed in said cover web.
18. The optical sensor assembly as set forth in claim 13, wherein
said plurality of sample chambers are disposed in fixed relation on
said optical sensor.
19. A method of operating an optical sensor, comprising the steps
of: (a) providing an optical sensor including: i) a substrate web
of predetermined length, the substrate web being substantially gas
impermeable and optically transparent in a predetermined spectral
range; ii) a plurality of elongated sensor stripes extending in
parallel, spaced relation along the length of said web, each one of
said plurality of sensor stripes adapted for providing an optically
discernible response to presence of at least one of a plurality of
discrete analytes; iii) said optical sensor adapted for selective
analyte-sensing contact with the plurality of samples, wherein each
one of the plurality of samples are selectively superimposable with
each one of said plurality of elongated sensor stripes at one of a
plurality of discrete sample positions along the lengths thereof;
iv) said optically discernible response being substantially
identical at said plurality of discrete sample positions along the
length thereof; v) wherein the plurality of samples comprises at
least one unknown sample and at least one calibration sample, the
optical sensor adapted for being calibrated upon disposition of the
calibration sample in said analyte-sensing contact with said
optical sensor at one of said discrete sample positions distinct
from that of said at least one unknown sample; (b) placing the
calibration sample in said analyte-sensing contact with the optical
sensor at one of said plurality of discrete sample positions along
the lengths of the sensor stripes; (c) measuring optical response
of the optical sensor at the one of the plurality of discrete
sample positions; (d) obtaining calibration data utilizing the
optical response of the one of the plurality of discrete sample
positions; (e) placing the at least one unknown sample in said
analyte-sensing contact with the optical sensor at another of the
plurality of discrete sample positions along the lengths of the
sensor stripes; (f) measuring optical response of the other of the
plurality of discrete sample positions; (g) utilizing the
calibration data obtained for the one of the plurality discrete
sample positions for calibration of the optical response of the
other of the plurality of discrete sample positions.
20. The method as set forth in claim 19, wherein said step of
utilizing (g) further comprises calculating presence and
concentration of an analyte disposed in the at least one unknown
sample.
21. The method as set forth in claim 19, wherein said steps of:
placing (b) and measuring (c) are undertaken substantially
simultaneously with said steps of placing (e) and measuring (f),
respectively.
22. The method as set forth in claim 19, wherein said step of
placing (e), further comprises the step of placing the at least one
unknown sample in said analyte-sensing contact with the optical
sensor adjacent the at least one of the plurality of discrete
sample positions.
23. The method as set forth in claim 19, wherein: said step of
placing (b) includes placing a calibration sample at a
predetermined number of the plurality of discrete sample positions
along the lengths of said sensor stripes; and said step of
obtaining (d) includes obtaining calibration data utilizing the
optical response of the predetermined number of the plurality of
discrete sample positions.
24. The method as set forth in claim 19, wherein: said step of
placing (b) includes placing a calibration sample at at least two
of the plurality of discrete sample positions along the lengths of
the sensor stripes, the two being disposed on opposite sides of the
other of the plurality of discrete sample positions along the
lengths of the sensor stripes; and said step of obtaining (d)
includes obtaining calibration data utilizing the optical response
of the two of the plurality of discrete sample positions along the
lengths of the sensor stripes for calibration of the optical
response of the other of the plurality of discrete sample positions
along the lengths of the sensor stripes.
25. The method as set forth in claim 19, further comprising the
steps of: placing other ones of the plurality of samples in
analyte-sensing contact with other ones of the plurality of
discrete sample positions along the lengths of the sensor stripes,
proximate the at least one of the plurality of discrete sample
positions; measuring optical response of the other ones of the
plurality of discrete sample positions; and utilizing the
calibration data obtained for the at least one discrete sample
position for calibration of the optical response of the other ones
of the plurality of discrete sample positions.
26. The method as set forth in claim 25, wherein: said step of
placing (b) includes placing a calibration sample at a
predetermined number of the plurality of discrete sample positions
along the lengths of said sensor stripes; and said step of
obtaining (d) includes obtaining calibration data utilizing the
optical response of the predetermined number of the plurality of
discrete sample positions.
27. The method as set forth in claim 19, wherein: said step of
providing (a) includes providing an optical sensor assembly
including the optical sensor, a plurality of sample chambers
superimposed in parallel, spaced relation on said web and being
superimposed with said plurality of elongated sensor stripes at a
plurality of discrete sample positions along the lengths thereof,
wherein each of the plurality of sample chambers is adapted for
alternately maintaining individual ones of the plurality of samples
in said analyte-sensing contact; said step of placing (b) includes
placing a calibration sample in a first one of the plurality of
sample chambers; said step of measuring (c) includes measuring
optical response of the optical sensor at the first one of the
plurality of sample chambers; said step of placing (e) includes
placing an unknown sample in a second one of the plurality of
sample chambers, the second one of the plurality of sample chambers
being disposed adjacent the first one of the plurality of sample
chambers; said step of measuring (f) includes measuring optical
response of the optical sensor at the second one of the plurality
of sample chambers; and said step of utilizing (g) includes
utilizing the calibration data obtained from the first one of the
plurality of sample chambers for calibration of the optical
response obtained from the second one of the plurality of sample
chambers disposed adjacent thereto.
28. The method as set forth in claim 27, wherein: said step of
placing (b) includes placing a calibration sample at a plurality of
first ones of the plurality of sample chambers; and said step of
obtaining (d) includes obtaining calibration data utilizing the
optical response of the plurality of first ones of the plurality of
sample chambers.
29. The method as set forth in claim 27, wherein: said step of
placing (b) includes placing a calibration sample at at least two
of a plurality of first ones of the plurality of sample chambers,
the at least two being disposed on opposite sides of a second one
of the plurality of sample chambers; and said step of obtaining (d)
includes obtaining calibration data utilizing the optical response
of the at least two of a plurality of first ones for calibration of
the optical response of the second one of the plurality of sample
chambers.
30. The method as set forth in claim 29, wherein said step of
placing (b) includes placing a calibration sample at at least two
sample chambers disposed on opposite sides and adjacent a second
one of the plurality of sample chambers.
31. The method as set forth in claim 27, wherein said steps of
placing (b) and measuring (c) are undertaken substantially
simultaneously with said steps of placing (e) and measuring (f),
respectively.
32. The method as set forth in claim 27, wherein: said step of
placing (b) further includes placing a calibration sample in each
of a plurality of first ones of the plurality of sample chambers;
said step of measuring (c) further includes measuring optical
response of the optical sensor at each of the plurality of first
ones of the plurality of sample chambers; said step of placing (e)
further includes placing an unknown sample in respective second
ones of the plurality of sample chambers, each of said second ones
of the plurality of sample chambers being disposed adjacent one of
said first ones of said plurality of sample chambers; said step of
measuring (f) further includes measuring optical response of the
optical sensor at each of the plurality of second ones of said
plurality of sample chambers; and said step of utilizing (g)
further includes utilizing the calibration data from each of the
first ones to analyze the optical response of the second ones of
the plurality of sample chambers disposed adjacent thereto.
33. The method as set forth in claim 32, wherein said steps of
placing (b) and measuring (c) are undertaken substantially
simultaneously, respectively, with the steps of placing (e) and
measuring (f), for each pair of adjacent first ones and second
ones.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] This invention relates to chemical analysis of liquids, and
more particularly, to an optical sensor for sensing analyte content
of biological fluids such as blood.
[0003] 2. Background Information
[0004] Chemical analysis of liquids, including biological fluids
such as blood, plasma or urine is often desirable or necessary.
Sensors that utilize various analytical elements to facilitate
liquid analysis are known. These sensing elements have often
included a reagent in either a wet or dry form sensitive to a
substance or characteristic under analysis, termed analyte herein.
The reagent, upon contacting a liquid sample containing the
analyte, effects formation of a colored material or another
detectable change in response to the presence of the analyte.
Examples of dry analytical sensing elements include pH test strips
and similar indicators wherein a paper or other highly absorbent
carrier is impregnated with a material, chemically reactive or
otherwise, that responds to contact with liquid containing hydrogen
ion or other analyte and either generates color or changes color.
Specific examples of such test strips are disclosed in European
publication No. EP 0119 861 B1, which describes a test for
bilirubin; in U.S. Pat. No. 5,462,858 which describes a dry
multilayer strip for measuring transaminase activity; and U.S. Pat.
No. 5,464,777 which discloses a reflectance based assay for
creatinine. While providing a convenience factor, in that they can
be stored dry and are ready to use on demand, these individual test
elements are generally utilized in "wet" blood or serum chemistry,
wherein the strips become saturated during use. This hydration and
the depletion of reactive chemical reagents effectively prevents
their re-use. This aspect also complicates handling and disposal of
the multitude of individual used test elements.
[0005] Alternatively, some analytes can be measured with a sensing
element which is used repeatedly after an initial wet-up and
calibration and with washes between samples. For example a
reuseable electrochemical sensor for oxygen is described in
commonly assigned U.S. Pat. No. 5,387,329 and a reuseable
electrochemical sensor for glucose is described in commonly
assigned U.S. Pat. No. 5,601,694. These sensors function within the
context of a complex piece of support instrumentation to perform
the repetitive calibration and wash functions.
[0006] Other analytical sensing elements which are based on an
optical signal response are disclosed in U.S. Pat. Nos. 4,752,115;
5,043,286; 5,453,248 and by Papkovsky et al in Anal. Chem. vol 67
pp 4112-4117 (1995) which describe an oxygen sensitive dye in a
polymer membrane, as does commonly assigned U.S. patent application
Ser. No. 08/617,714, which is hereby incorporated in its entirety,
herein. Examples of an optical CO.sub.2 sensor are described in
U.S. Pat. Nos. 4,824,789; 5,326,531 and 5,506,148. These elements
utilize a polymer based membrane chemistry to achieve advantages in
storage, and continuous use or re-use as compared to the wetable or
hydrated single use chemistry strips. Analytical elements of this
type are typically adapted for multiple uses within a single sample
chamber of an optical sensor assembly. In operation, a fluid sample
of unknown analyte content (an "unknown sample") is tested by
inserting the sample into the sample chamber where it contacts the
analytical element. A change in the optical properties of the
analytical element is observed. Such an observation is then
compared to calibration data previously obtained by similarly
testing a calibration liquid of known analyte content. In this
manner, characteristics of the analyte of interest in the unknown
sample are determined.
[0007] An example of a single use optical sensor application of
this normally reuseable type is known as a "AVL OPTI 1" available
from AVL List GmbH of Graz, Austria. While sensors of this type may
operate satisfactorily in many applications, they are not without
limitations. In particular, they rely on sequential steps for
calibration and subsequent sample readings, in which each such
sensing device must be individually calibrated prior to testing an
unknown sample. This technique is required due to variations in
analytical elements from sensor to sensor. These variations may be
attributed to a variety of factors, including manufacturing
variables such as differences in individual lots, and distinct
storage histories.
[0008] Sequential calibration and sample reading may
problematically lead to sample contamination in the event the
sample chamber and analytical elements are insufficiently washed
between samples. In addition, the calibration is time consuming and
may delay analysis of the unknown sample. This delay may be
particularly inconvenient in some operating environments such as,
for example, critical care facilities.
[0009] An additional disadvantage of the sequential approach is the
temporal variation or time delay between testing of the calibrant
and testing of the unknown sample. This variation may provide a
potential opportunity for inaccuracies in test results.
[0010] Further, discarded wash fluid comprises approximately 80% of
the waste generated by such conventional sensor based testing
techniques. This waste is classified as biohazardous particularly
if it is co-mingled with biological samples and thus disposal
thereof is relatively expensive, both in economic and environmental
terms. This waste also poses a potential health risk to health care
workers and those who may otherwise come into contact with the
waste during or after disposal.
[0011] Thus, a need exists for an improved optical sensor that
eliminates the need for serial calibration and addresses the
problems of waste generation inherent in sensor practices of the
prior art while retaining the advantages of disposable, use on
demand, devices.
SUMMARY OF THE INVENTION
[0012] According to an embodiment of the present invention, an
optical sensor adapted for sensing analyte content of a plurality
of samples is provided. The optical sensor comprises:
[0013] a substrate web of predetermined length, the substrate web
being substantially gas impermeable and optically transparent in a
predetermined spectral range;
[0014] a plurality of elongated sensor stripes extending in a
parallel spaced relation along the length of the web;
[0015] each one of the plurality of sensor stripes adapted for
providing an optically discernible response to presence of at least
one analyte;
[0016] the optical sensor adapted for selective analyte-sensing
contact with the plurality of samples, wherein each one of the
plurality of samples are selectively superimposable with each one
of the plurality of elongated sensor stripes at one of a plurality
of discrete positions along the lengths thereof;
[0017] the optically discernible response being substantially
identical at a plurality of discrete positions along the length
thereof.
[0018] In a first variation of this aspect of the present
invention, an optical sensor assembly adapted for sensing analyte
content of a plurality of samples is provided. The optical sensor
assembly comprises:
[0019] the optical sensor as set forth in the above-referenced
first aspect of the present invention;
[0020] at least one sample chamber selectively superimposable with
each of the plurality of elongated sensor stripes at the plurality
of discrete positions along the lengths thereof;
[0021] wherein the at least one sample chamber is adapted for
alternately maintaining individual ones of the plurality of samples
in the analyte-sensing contact.
[0022] In a second variation of the first aspect of the present
invention, an optical sensor assembly adapted for sensing analyte
content of a plurality of samples is provided. The optical sensor
assembly includes:
[0023] the optical sensor as set forth in the above-referenced
first aspect of the present invention;
[0024] a plurality of sample chambers disposed in parallel, spaced
relation on the web, each one of the plurality of sample chambers
being sealably superposed with the plurality of elongated sensor
stripes at one of a plurality of discrete positions along the
lengths thereof;
[0025] wherein each of the plurality of sample chambers is adapted
for alternately maintaining individual ones of the plurality of
samples in the analyte-sensing contact.
[0026] In a second aspect of the present invention, a method of
operating an optical sensor comprises the steps of:
[0027] (a) providing an optical sensor including:
[0028] i) a substrate web of predetermined length, the substrate
web being substantially gas impermeable and optically transparent
in a predetermined spectral range;
[0029] ii) a plurality of elongated sensor stripes extending in
parallel, spaced relation along the length of the web, each one of
the plurality of sensor stripes adapted for providing an optically
discernible response to presence of at least one of a plurality of
discrete analytes;
[0030] iii) the optical sensor adapted for selective
analyte-sensing contact with the plurality of samples, wherein each
one of the plurality of samples are selectively superimposable with
each one of the plurality of elongated sensor stripes at one of a
plurality of discrete positions along the lengths thereof;
[0031] iv) the optically discernible response being substantially
identical at a plurality of discrete positions along the length
thereof;
[0032] v) wherein the plurality of samples comprises at least one
unknown sample and at least one calibration sample, the optical
sensor adapted for being calibrated upon disposition of the
calibration sample in the analyte-sensing contact with the optical
sensor at a discrete position along the length of the sensor
stripes distinct from that of the at least one unknown sample;
[0033] (b) placing the calibration sample in the analyte-sensing
contact with the optical sensor at one of the plurality of discrete
positions along the lengths of the sensor stripes;
[0034] (c) measuring optical response of the optical sensor at the
one of the plurality of discrete positions;
[0035] (d) obtaining calibration data utilizing the optical
response of the one of the plurality of discrete positions;
[0036] (e) placing the at least one unknown sample in the
analyte-sensing contact with the optical sensor at an other of the
plurality of discrete positions along the lengths of the sensor
stripes;
[0037] (f) measuring optical response of the other of the plurality
of discrete positions;
[0038] (g) utilizing the calibration data obtained for the one of
the plurality discrete positions for calibration of the optical
response of the other of the plurality of discrete positions.
[0039] The above and other features and advantages of this
invention will be more readily apparent from a reading of the
following detailed description of various aspects of the invention
taken in conjunction with the accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0040] FIG. 1 is a plan view of an optical sensor of the present
invention;
[0041] FIG. 2 is a perspective view of an embodiment of an optical
sensor assembly of the present invention, including the optical
sensor of FIG. 1 and a sample chamber disposed thereon;
[0042] FIG. 3 is a cross-sectional elevational view taken along
FIG. 3-3 of FIG. 2;
[0043] FIG. 4A is a perspective view, with portions thereof peeled
back, of an alternate embodiment of an optical sensor assembly of
the present invention, including the optical sensor of FIG. 1 and a
plurality of sample chambers disposed thereon;
[0044] FIG. 4B is a view similar to FIG. 4A, of another alternate
embodiment of an optical sensor assembly of the present
invention;
[0045] FIG. 4C is a view similar to FIGS. 4A and 4B, of a further
alternate embodiment of an optical sensor assembly of the present
invention;
[0046] FIG. 5 is a schematic representation of a portion of a test
apparatus capable of use in combination with an optical sensor of
the present invention;
[0047] FIG. 6 is a schematic representation of a test apparatus
including the portion thereof shown in FIG. 5, capable of measuring
the output signal of a luminescent optical sensor of the present
invention;
[0048] FIG. 7A is a graphical representation of optical response of
a portion of an optical oxygen sensor of the type shown in FIGS. 1
and 4;
[0049] FIG. 7B is a graphical representation of response to aqueous
buffer samples, of the portion of the optical oxygen sensor
utilized to generate FIG. 7A;
[0050] FIG. 8 is a graphical representation of the response of an
optical oxygen sensor of the type shown in FIGS. 1 and 4 and
constructed from a second different membrane and dye
formulation;
[0051] FIG. 9 is a response curve similar to that of FIG. 7B, for a
carbon dioxide sensing portion of an optical sensor of the type
shown in FIGS. 1 and 4;
[0052] FIG. 10 is a graphical representation of response to
acidification of the fluorescein dye, of the portion of the optical
pH sensor described in FIGS. 1 and 4;
[0053] FIG. 11 is a graphical representation of the simultaneous
response of sensors of the present invention, for three analytes,
for three different known samples; and
[0054] FIG. 12 is a graphical response curve for a single oxygen
sensor stripe of the present invention calibrated by the use of
several known samples similar to those utilized to generate FIG.
11.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0055] Referring to the figures set forth in the accompanying
drawings, illustrative embodiments of the present invention will be
described in detail hereinbelow. For clarity of exposition, like
features shown in the accompanying drawings shall be indicated with
like reference numerals and similar features shown, for example, in
alternate embodiments in the drawings, shall be indicated with
similar reference numerals.
[0056] Briefly described, the present invention includes a multiple
single use optical sensor fabricated as a series of continuous
sensor stripes 14 deposited on a substrate web 12 (FIG. 1). One
sample chamber 16 (FIG. 2) or multiple sample chambers 116 (FIG. 4)
are adapted to extend transversely across a discrete portion of the
series of sensor stripes 14 to facilitate analysis of a sample
disposed therein. Sample chamber 16 may be moved, or additional
sample chambers utilized to enable subsequent measurements of
additional samples at unused discrete portions of sensor stripes
14. The continuous nature of the sensor stripes provides
consistency along the lengths thereof to enable calibration data
obtained from one discrete portion of a sensor stripe 14 to be
utilized for testing and determining presence and concentration of
analytes in an unknown sample disposed at an other discrete portion
of the sensor stripe. This aspect advantageously eliminates the
need for any particular discrete portion of a sensor stripe 14 to
be contacted by more than one sample for improved sensor
performance and reduced waste.
[0057] Throughout this disclosure, the term "analyte" shall refer
to any substance, compound, or characteristic such as, for example,
pH, capable of detection and/or measurement relative to a liquid
sample. Similarly, the term "concentration" shall refer to the
level or degree to which an analyte is present in a sample. The
term "axial" or "longitudinal" when used in reference to an element
of the present invention, shall refer to the relatively long
dimension or length thereof. For example, when used in connection
with an optical sensor of the present invention, "longitudinal"
shall refer to a direction substantially parallel to sensor stripes
14 thereof. Similarly, the term "transverse" shall refer to a
direction substantially orthogonal to the axial or longitudinal
direction. Moreover, the use of the term "calibration" or
"calibration sample" shall be understood to encompass a sample of
substantially any known analyte composition, including "QC" or
"quality control" samples commonly used by those skilled in the art
to help ensure uniformity between tests.
[0058] Referring now to the drawings in detail, as shown in FIG. 1,
an optical sensor 10 of the present invention includes a backing or
substrate web 12, with a plurality of sensor stripes 14 extending
longitudinally in parallel, spaced relation thereon. Backing web 12
is fabricated as a sheet from a material optically transparent in a
predetermined optical spectrum, as will be discussed hereinafter.
The backing web is preferably fabricated from a substantially
liquid and gas impermeable material, such as, for example, glass or
a thermoplastic material such as polyethylene terephthalate or
SARAN.RTM..
[0059] In this regard, those skilled in the art will recognize that
fabrication of the substrate web from relatively gas permeable
materials, such as, for example, Polytetrafluoroethylene (PTFE),
may disadvantageously distort analyte analysis. This is due to the
tendency for analytes to diffuse out of the sample, or for ambient
gases such as atmospheric Oxygen (O.sub.2) and/or Carbon Dioxide
(CO.sub.2), to leach out of the substrate and into the sensor
material and sample, during analysis. In a preferred embodiment,
substrate web 12 is fabricated as a film of polymeric plastic
material sold under the Dupont trademark Mylar.RTM.. Webs were
obtained from ERA Industries INC. in Seabrook N.H. In addition to
being substantially gas impermeable, this material advantageously
provides substrate web 12 with flexibility, as will be discussed in
greater detail hereinafter. The substrate web may be fabricated
using any convenient method common in the art, such as conventional
molding, casting, extrusion or other suitable thin-film fabrication
techniques.
[0060] Each sensor stripe 14 may be fabricated as a series of
discrete portions, such as a series of dots, arranged in a row
extending longitudinally along the substrate web. Alternatively, in
a preferred embodiment as shown, each sensor stripe 14 extends
substantially continuously in the longitudinal direction. Each
sensor stripe 14 comprises at least one of any number of analytical
elements, including substances, compounds or structures known to
those skilled in the art to be optically sensitive to a
predetermined analyte. Such optical sensitivity may include, for
example, exhibition of optically discernible changes in
reflectance, refractive index, light transmittance, or in a
preferred embodiment, luminescence, which may encompass emitted
light in the form of either phosphorescence or fluorescence.
[0061] Examples of analytes that may be analyzed include BUN (blood
urea nitrogen), glucose, calcium (Ca.sup.++), potassium (K.sup.+),
sodium (Na.sup.+), pH, and partial pressures of carbon dioxide
(pCO.sub.2) and oxygen (PO.sub.2). Preferred analytical elements
include, for example, analytical elements for carbon dioxide
(pCO.sub.2) as disclosed in U.S. Pat. Nos. 5,387,525 (the '525
patent) and 5,506,148 (the '148 patent), an analytical element for
pH as disclosed in International Publication No. WO 95/30148 and by
Bruno, et al. in Anal. Chem. Vol 69, pp. 507-513 (1997) and an
analytical element for oxygen (pO2) as disclosed in U.S. patent
application Ser. No. 08/617,714, all of which are hereby
incorporated by reference in their entireties, herein. All of these
preferred analytical elements emit characteristic luminescence
which is responsive to the presence of their respective analytes
when subjected to incident light of a predetermined spectral
wavelength or spectral range.
[0062] In a preferred embodiment, each sensor stripe 14 comprises a
single analytical element. However, it is contemplated that a
single sensor stripe of the present invention may comprise a
plurality of analytical elements, each of the plurality of
analytical elements exhibiting an independently measurable response
to presence of their respective analytes. In this regard, for
example, a single sensor stripe 14 may comprise first, second and
third analytical elements. The first analytical element may exhibit
enhanced fluorescence in presence of a first analyte when subjected
to incident light in a first spectral range. The second analytical
element may exhibit diminished phosphorescence in presence of a
second analyte when subjected to incident light in a second
spectral range. The third analytical element may, for example,
exhibit another optical response, such as enhanced reflectance, in
presence of a third analyte when subjected to incident light in a
predetermined spectral range.
[0063] Sensor stripes 14 are applied to the substrate web 12 by any
convenient means, either by batch or continuous processes. For
example, stripes 14 may be applied by conventional printing
techniques, such as silk screen or other lithographic techniques.
It is also contemplated that laser or ink jet printing technologies
may ultimately be adapted for application of the sensor stripes.
Alternatively, the stripes may be applied by continuous direct
deposition or painting-type application techniques as well as by
spray painting.
[0064] For example, in a preferred embodiment, one may use a micro
dispensing system of the type commercially available from Gilson,
Worthington, Ohio; Cavro Scientific Instruments Inc., Sunnyvale,
Calif.; Elder Laboratories Inc., Napa, Calif.; IVEK Corp.,
Springfield, Vt.; or Fluid Metering Inc., Oyster bay, N.Y., as well
as other commercial sources for chromatographic delivery systems.
Operation of this equipment is familiar to those of skill in the
art. Briefly described, the material comprising the sensor stripe,
including at least one analytical element, is prepared in liquid
form and fed to a nozzle of predetermined size and shape, suspended
or superposed over substrate web 12. The liquid is expressed from
the nozzle at a predetermined rate onto the substrate web as the
web is moved longitudinally at a predetermined rate relative the
nozzle with either reciprocating or rolled web technologies of a
more continuous nature. This process is repeated at spaced
locations along the transverse dimension or width of the substrate
web for each sensor stripe. The liquid is then dryed or cured in a
conventional manner to form a solid sensor stripe 14.
[0065] While the aforementioned method for deposition of sensor
stripes 14 is preferred, substantially any method of deposition may
be utilized that enables the mechanical and optical properties of
sensor stripes 14 to be held substantially constant over the
lengths thereof. In this regard, parameters such as stripe
thickness, width, contour, and composition are maintained at
predetermined levels to provide sensor response that is relatively
constant or identical at various positions along the length of each
sensor stripe 14. Moreover, the skilled artisan will recognize that
sensor response will be particularly consistent over relatively
short sections of the stripe. In other words, the uniformity of
response of discrete portions of a sensor stripe 14 will be in some
measure proportional to the spatial distance therebetween.
[0066] Referring now to FIG. 2, an optical sensor assembly 15 of
the present invention includes a sample chamber 16 adapted for use
in combination with optical sensor 10. Sample chamber 16 comprises
an elongated, substantially tubular cavity 18 disposed within an
elongated chamber member 19. Cavity 18 has a transverse
cross-section nominally uniform along the length thereof and
defined, in part, by a substantially concave or recessed surface
21, best shown in FIG. 3. Throughout this disclosure, the term
"concave" shall refer to any substantially hollowed out recess or
cavity, regardless of whether the surface thereof is curved or
comprises a plurality of substantially flat surfaces as shown
herein. In this regard, referring to FIG. 3, concave surface 21
extends inwardly from a substantially planar engagement surface 24
of chamber member 19.
[0067] As shown in FIGS. 2 and 3, engagement surface 24 is adapted
for being superimposed transversely across, preferably in slidable,
surface-to-surface engagement with substrate web 12 and sensor
stripes 14. So disposed, a discrete portion of web 12, including
portions of sensor stripes 14, effectively closes concave surface
21, to thus define a longitudinal side wall of tubular cavity 18.
Moreover, engagement surface 24, substrate web 12 and sensor
stripes 14 are each sufficiently smooth that upon application of a
predetermined force tending to maintain such surface-to-surface
contact, a fluid-tight seal is maintained therebetween. Sample
chamber 16 is thus adapted for supportably maintaining a fluid
sample in surface to surface or analyte-sensing contact with a
discrete portion of each sensor stripe 14, as will be discussed in
greater detail hereinafter with respect to operation of the
embodiments of the present invention.
[0068] As shown in FIG. 2, entry and exit apertures 20 and 22,
respectively, each extend through chamber member 19. The apertures
each extend orthogonally to, and in communication with, cavity 18
at opposite ends thereof, to facilitate sample flow into and out of
sample chamber 16.
[0069] As shown, sample chamber 16 is a reusable device, adapted
for either multiple tests at a particular discrete location on
sensor stripes 14, or alternatively, progressive movement to fresh
(unused) portions of the sensor stripes for successive sample
testing. These alternative testing techniques will be discussed
hereinafter with respect to operation of the present invention.
[0070] Referring now to FIG. 4A, an alternate embodiment of the
present invention is shown as optical sensor assembly 115. This
optical sensor assembly includes multiple individual sample
chambers 116 disposed on optical sensor 10. Sensor assembly 115 is
preferably fabricated as a laminate comprising optical sensor 10,
an intermediate or chamber web 26 and a cover web 28.
[0071] Chamber web 26, in combination with cover web 28, comprise
sample chambers 116. As shown, chamber web 26 is an elongated sheet
that includes a series of transversely extending cavities 118. The
cavities are spaced at predetermined distances from one another
along the length of the web. Web 26 is preferably fabricated from a
material and in a manner similar to that of substrate web 10.
Cavities 118 are formed by any convenient method, such as, for
example, by subjecting web 26 to conventional die-cutting
operations. Alternatively, in the event web 26 is fabricated by
molding, cavities 118 may be molded integrally therewith.
[0072] Cover web 28 is superimposed or laminated in a sealed,
fluid-tight manner over chamber web 26. This combination of chamber
web 26 and cover web 28 effectively provides each chamber 116 with
a transverse cross-section defined by concave surface 21 as
described hereinabove with respect to FIG. 3. A series of entry and
exit bores or apertures 20 and 22 extend through cover web 28 in
communication with opposite ends of cavities 118 as also discussed
hereinabove. Alternatively, the bores or apertures 20 and 22 may
also be formed in the substrate web itself 12 or used in
combination with apertures in the cover web 28. Cover web 28 is
preferably fabricated from a material and in a manner similar to
that of both substrate web 12 and chamber web 26. Any conventional
means, including, for example, ultrasonic and vibration welding or
adhesives of various types may be utilized to laminate cover web 28
to chamber web 26. In a preferred embodiment, however, a
conventional adhesive is utilized to bond webs 26 and 28 to one
another.
[0073] Chamber web 26 is laminated onto optical sensor 10 so that
sensor 10 effectively closes and seals concave surfaces 21 of each
cavity 118 in a manner similar to that described hereinabove with
respect to cavity 18. Thus, rather than being movable as is cavity
18 described hereinabove, cavities 118 are preferably immovably or
permanently disposed at spaced intervals along the length of
optical sensor 10. The manner in which chamber web 26 is laminated
onto optical sensor 10 is similar to that in which chamber web 26
is bonded to cover web 28.
[0074] Turning now to FIG. 4B, a further alternate embodiment is
shown as optical sensor assembly 115'. Assembly 115' is
substantially similar to optical sensor assembly 115, with the
distinction that entry and exit apurtures 20' and 22' are disposed
in substrate web 12, rather than in web 28.
[0075] An additional, similar alternative embodiment is shown in
FIG. 4C as optical sensor assembly 115". In assembly 115", some of
the entry and exit apertures (i.e. exit apertures 22 as shown) are
disposed in web 28 while others of the entry and exit apertures
(i.e. entry apertures 20') are disposed in substrate web 12.
[0076] Preferred embodiments of the invention having been
described, the following is a description of the operation thereof.
Referring initially to optical sensor assembly 15, as shown in
FIGS. 2 and 3, a sample to be tested is inserted into entry
aperture 20, such as by a pump means (not shown but which may
include the use of capillary forces or negative or postive
pressures). The sample is inserted until it substantially fills
sample chamber 16 and is thus placed in analyte-sensing contact
with a discrete portion of each respective sensor strip 14 as
discussed hereinabove. Once so disposed, any of a variety of
suitable instruments may be utilized to measure optical response of
the discrete portions to determine the existence and/or
concentration of analytes in the sample. Examples of such
instrumentation include a commercially available fluorimetric
device known as a model LS50-b Spectrofluorimeter available from
Perkin Elmer Corporation of Norwalk, Conn. A solid sample holder
accessory was specifically modified to accept the striped film
sensors for front face fluorescence measurements. By "front face"
or "front surface" it is meant that excitation and emission
collection is off the same surface. Illumination and collection
optics permit transmission of the excitation and emission signals
through the Mylar.RTM. substrate. Samples were introduced into a
hollowed out aluminum sample chamber located on the side of the
Mylar.RTM. opposite from the illumination and collection optics and
with the opening covered by the sensor stripe so that samples
contacted the stripe directly. Sample measurements with this device
are provided in Example 6 (FIG. 9) and Example 8 (FIG. 10).
[0077] Alternatively, a test apparatus 140 as depicted in FIG. 5
may be utilized. Briefly described, such an apparatus 140 includes
a flow cell assembly 60 and an excitation source and detector
sub-system 100 such as that disclosed in U.S. patent application
Ser. No. 08/617,714, and which is incorporated by reference in its
entirety herein. Sub-system 100 emits a beam of light having a
predetermined wavelength or spectral range. The light is directed
through fiber optic cable 80 onto the surface of substrate web 10
directly opposite a stripe 14 in sample chamber 16. The light
passes through the web, which, as mentioned hereinabove, is
substantially transparent thereto, wherein the light is incident on
a predetermined one of the sensor stripes 14. The incident light
serves to excite a portion of sensor stripe 14. Stripe 14 then
exhibits an optical response that corresponds to parameters (e.g.
presence and/or concentration) of the predetermined analyte in the
sample disposed in the sample chamber. This optical response is
received by detector sub-system 100.
[0078] The calibration information for the optical sensor assembly
is obtained by inserting a calibration sample or calibrant of known
analyte composition into the sample chamber and measuring response
of the sensor stripes thereto, in a manner substantially similar to
testing an unknown sample.
[0079] Referring now to FIGS. 5 and 6, test apparatus components 60
and 100 are described in additional detail. As shown in FIG. 5,
flow cell assembly 60 is adapted to receive an optical sensor 10
for measurement. Radiation or light impinging upon substrate web 12
and emitted from stripe 14 is respectively guided to and from
source and detection sub-system 100 by a fiber optic cable 80.
Cable 80 includes a core 82, cladding 84 and sheath 86 where the
core 82 and cladding 84 may be constructed from either glass or
plastic polymer materials. Cable 80 is imbedded into base 62 which
preferably has a low permeability to gases and a flat surface for
contact with substrate 12. Base 62 may comprise stainless steel or
another hard, thermally conductive material which is capable of
assisting in controlling the temperature of membrane 14. Source
radiation from cable 80 passes through substrate 12 and excites the
luminescent dye molecules dispersed within membrane 14. Elongated
member 19, including sample chamber 16, is pressed flat against
optical sensor 10 as discussed hereinabove. Alternatively, optical
sensor assembly 115 (FIG. 4), including sample chambers 116 (FIG.
4) may be utilized. Samples may be entered and subsequently removed
through the entrance and exit apertures 20 and 22. The signal from
each individual stripe 14 is then transmitted by cable 80 and
returned to source and detector sub-system 100.
[0080] Referring to FIG. 6, the measurement apparatus 140 is
comprised of flow cell assembly 60 and source and detector
subsystem 100. For the optical source and detector sub-system 100
an LED source 152, and lens 154 are used to launch excitation light
through filter 162 into one leg 182 of the fiber optic splitter 180
(avilable from American Laubscher Corp., Farmingdale, N.Y.). The
luminescent or emitted light signal returning from the sensor 10
down fiber cable 80 and leg 184 is passed through filter 168 and
aperture 158 before detection by photodiode 172. The output current
of emission detector 172 is amplified with a preamplifier 174, such
as a Stanford Research SR570 current preamplifier, converted to a
voltage and recorded for use in analysis. For example, with the pH
sensing dye fluorescein used in a sensor stripe, a
Panasonic.RTM.Blue LED (P389ND available from Digikey, Theif River
Falls, Minn.) would be preferred for source 152. A 485 nm center
wavelength 22 nm half bandwidth filter (available from Omega
Optical, Brattleboro, Vt.) would be preferred for filter 162 and a
535 nm center wavelength 35 nm half bandwidth filter, also
available from Omega Optical, Brattleboro, Vt. would be preferred
for filter 168. It should also be evident that each individual
sensor stripe, employing a different dye, will require its own
preferred LED source 152, excitation interference filter 162 and
emission interference filter 168. While particular arrangements of
optical source and detection systems have been disclosed herein,
other equivalent instruments are known to those skilled in the art
and are intended to be within the scope of the present
invention.
[0081] Testing procedures are undertaken at each sensor stripe 14
in sample chamber 16, either sequentially or in parallel, to test
for all of the predetermined analytes. Once analysis is complete,
the pump means removes the sample from chamber 16 through exit
aperture 22.
[0082] Analysis of subsequent samples, as well as the
aforementioned analysis of a calibration sample, may be
accomplished in a manner common to prior art sensors. Namely,
sample chamber 16 may be flushed with wash fluid to remove traces
of the previous sample from the sample chamber and sensor stripes.
Sample chamber 16 and the same discrete portions of sample stripes
14 with which the sample chamber is superposed, may be re-used for
a subsequent test sample. In this manner, sensor assembly 15 may
function as a conventional `multiple use` device. Alternatively the
present invention includes use of optical sensor 10 as a `multiple
single use device` in which subsequent tests may be performed at
discrete unused portions of sensor stripes 14. In this regard,
after testing is completed, sample chamber 16 may be washed and
dried sufficiently to clear any sample traces from chamber member
19 and prevent liquid carryover to the next chosen position. Sample
chamber 16 may then be moved relative the length of optical sensor
10 to superimpose cavity 18 with an unused portion of sensor
stripes 14. Once so disposed, a subsequent sample may be fed into
sample chamber 16 for analyte analysis. These steps may be
reiterated, so that a fresh discrete portion of each sensor stripe
14 is used for each sample (calibrant or unknown) in either a
sequential or simultaneous manner.
[0083] However, the present invention is preferably used in the
`multiple single use` mode when it is combined with provisions for
a plurality of sample chambers 116, as shown in FIG. 4, to enable
each sample chamber to be used only once. This nominally eliminates
the need for washing operations and each sample chamber effectively
becomes a waste container for its own sample. In addition, this
aspect substantially eliminates the potential for
cross-contamination of samples occasioned by repeated use of sample
chambers, as mentioned hereinabove.
[0084] An additional advantage of this construction is the ability
to conduct parallel testing of unknown and calibration samples. In
this regard, sample chambers 116 disposed proximate, and preferably
adjacent, one another may be utilized for simultaneously testing
calibration samples and unknown samples. Such parallel,
simultaneous testing provides additional precision in testing not
available with prior art devices by effectively eliminating any
inaccuracies in sensor response occasioned by temporal variations
between tests of calibration and unknown samples.
[0085] Moreover, in a further variation, both sensor assembly 15
(FIG. 2) and sensor assembly 115 (FIG. 4) may be calibrated at
multiple discrete positions along the lengths of sensor stripes 14.
This advantageously provides additional data points for increased
precision of the calibration information. In this regard, for still
further precision, calibration samples may be tested in chambers
disposed on opposite sides of, and adjacent to, a sample chamber
containing an unknown sample.
[0086] This multiple position calibration also facilitates
utilization of discrete calibration samples having different
combinations of analytes disposed therein. This aspect tends to
enhance the stability of the individual calibration mixtures by
enabling separation of analytes, such as, for example, glucose and
oxygen. One skilled in the art will recognize that the presence of
oxygen in a glucose solution tends to favor oxidative microorganism
growth. Thus, it is advantageous to have separate oxygen and
glucose calibration solutions. In general, a first calibration
sample may be provided with a first predetermined combination of
analytes, and a second calibration sample provided with a second
predetermined combination of analytes. The first and second
calibration samples then may be tested simultaneously at discrete
positions of sensor stripes 14. The data obtained from testing
these separate calibration samples may be combined for analyzing
test results for unknown samples at the same or other discrete
positions along sensor stripes 14.
[0087] Thus, as discussed hereinabove, rather than rely on temporal
stability, the present invention relies on spatial stability,
namely the assumption that sensor portions located proximate one
another along the sensor stripes will exhibit substantially
identical response characteristics. This reliance is made possible
by the deposition of the analytical elements as substantially
continuous sensor stripes 14 as discussed hereinabove, with
increased precision enabled, as desired, through the use of
adjacent sample chambers 116 for respective testing and
calibration.
[0088] Moreover, the combination of spatial and temporal proximity
in these measurements permits the use of conventional differential
and ratiometric techniques to further improve accuracy and
precision thereof. In particular, by introducing and measuring an
unknown sample and a calibrant into respective sample chambers at
the same time, it is possible to simultaneously observe and compare
the response dynamics of the calibrant versus the unknown sample to
further enhance accuracy of response measurement.
[0089] The construction of the present invention also addresses the
problem of storage history variations that tend to compromise
performance and consistency of prior art sensors. For example,
otherwise identical prior art sensors may have been stored for
different periods of time or exposed to variations in environmental
conditions (e.g. differences in temperature, humidity or radiation)
during storage, that may impact consistency between sensors. By
virtue of fabricating the analytical elements as nominally
continuous stripes on a single substrate, the present invention
tends to ensure that each discrete portion of sensor stripes 14 has
an identical storage history to further improve sensor
consistency.
[0090] Moreover, the present invention, particularly sensor
assembly 115, provides an additional advantage in terms of waste
reduction. As mentioned hereinabove, approximately 80% of waste in
connection with prior art sensors comprises wash fluid used to
clean the sample chamber and analytical elements between unknown
samples. Such waste is generally classified as biohazardous, thus
requiring relatively rigorous and expensive special handling. By
substantially reducing or eliminating the washing requirements
through the construction of individualized sample chambers 116 as
discussed hereinabove, the present invention effectively reduces
biohazardous waste relative to prior art devices, for desireable
cost and safety improvements.
[0091] The following illustrative examples are intended to
demonstrate certain aspects of the present invention. It is to be
understood that these examples should not be construed as limiting.
In the examples, sensor stripes 14 were deposited on a 75
micrometers (m) thick Mylar.RTM. substrate web 12 positioned with
an IVEK LS Table. Deposition of the polymer and dye formulations
was achieved with a micro dispensing system of the type discussed
hereinabove. Examples of the construction of striped sensor
membranes and demonstrations of their functionality are given in
the following:
EXAMPLE 1
[0092] Into one ml of the solvent tetrahydrafuran (THF) from Alrich
(Milwaukee, Wis.) were dissolved 100 mg of polystyrene (MW=280,000
and obtained from Scientific Polymer Products Inc. in Ontario,
N.Y.) and 2 mg of the oxygen sensing dye octaethyl-Pt-porphyrin
ketone (OEPK) purchased from the Joanneum Research Institute in
Graz Austria. The viscosity of the solution was 37 centipoise (cps)
as measured on a Brookfield RVDVIIIC/P Rheometer. The mixture was
then deposited through a nozzle located 75 m above a clear
Mylar.RTM. film and at a rate of 5 ml/sec with a Digispense 2000
pump system from IVEK to produce a stripe at a linear rate of 50
mm/sec, having a width of approximately 2 mm and a thickness of
about 5 m when dried. After air drying, the stripes were cured at
11.degree. C. for one hour under a vacuum and cooled to remove all
traces of solvent. The resultant oxygen sensing stripes were
translucent and of a light purple color.
EXAMPLE 2
[0093] A sensor stripe from example 1 was placed in the measurement
device described with respect to FIG. 5 but altered to contain the
appropriate yellow LED source, an Omega 585DF20 excitation filter,
and a Omega 750DF50 emission filter for the dye
octaethyl-Pt-porphyrin ketone. A flowing gas stream with differing
partial pressures of oxygen corresponding to 0%, 100%, 26%, 12%,
7%, 12%, 26%, 100% and finally 0% oxygen was passed over the sensor
and the luminescence elicited from the dye recorded. The
luminescence quenching trace in FIG. 7A was used to derive a
Stem-Volmer quenching constant of 0.026 (mmHg).sup.-1. The exposure
of the striped oxygen sensing membrane to duplicate aqueous buffer
samples tonometered to partial pressures of 92, 43 and 171 mm Hg
oxygen also produced rapid, and reversible responses as documented
in FIG. 7B which could be used to quantitate the amount of
dissolved oxygen in solution.
EXAMPLE 3
[0094] A sensing stripe for the analyte oxygen was constructed as
follows. The dye octaethyl-Pt-porphyrin was synthesized according
to methods described in J. Molecular Spectroscopy 35:3 p359-375
(1970). The styrene/acrylonitrile copolymer, with MW=165,000 and
containing 25% acrylonitrile, was obtained from Scientific Polymer
Products Inc., Ontario, N.Y. A mixture of 2 mg dye and 100 mg of
copolymer dissolved into 1 ml of THF was deposited on a Mylar.RTM.
film as in example
EXAMPLE 4
[0095] A sensor stripe from example 3 was placed in the measurement
device described hereinabove with respect to FIG. 5 and a flowing
gas stream with differing partial pressures of oxygen corresponding
to 0%, 26% and finally 100% oxygen were passed over the sensor. The
luminescence elicited with green 540 nm excitation light from the
octaethyl-Pt-porphyrin dye was continuously measured at 650 nm and
the luminescence quenching trace recorded as shown in FIG. 8.
EXAMPLE 5
[0096] An analytical element for CO.sub.2 was fabricated
substantially as set forth in the above-referenced '525 and '148
patents. Namely, a 7% solution (by weight) of ethyl cellulose was
prepared by dissolving 7 g in 100 ml of a 7:3 toluene:ethanol
mixture. To this solution was added 5 mg of hydroxpyrenetrisulponic
acid (HPTS). 2 ml of Tetrabutylamonium hydroxide was added to the
mixture. The solution striped at a linear rate of 50 mm/sec with a
solution delivery rate of 5 ml/sec with the nozzle located 75 m
above the substrate. After air drying overnight this produced very
faintly green stripes for CO.sub.2 sensing.
EXAMPLE 6
[0097] A portion of the striped CO.sub.2 sensor in example 5 was
placed in an optical chamber on a Perkin Elmer LS-50B
spectrofluorimeter. Front surface illumination and collection
optics permitted transmission of the 460 nm excitation and 506 nm
emission signals through the Mylar.RTM. substrate. Tonometered
liquid samples were introduced into a hollowed out aluminum sample
chamber with an opening covered by the sensor stripe. Introduction
of increasing partial pressures of CO.sub.2 corresponding to 5.66
and 8.33% CO.sub.2 caused reversible fluorescence changes as
documented in FIG. 9.
EXAMPLE 7
[0098] Fifty mg of a pH sensitive copolymer composed of
N,N-Dimethylacrylamide and N-tert-butylacrylamide monomers with a
covalently linked 4-acrylamidofluorescein was dissolved into 1 ml
of THF in the manner described by Alder et al. in the
above-referenced patent application WO 95/30148. The polymer
solution was striped at a speed of 50 mm/sec and dispensed at a
rate of 4 ml/sec from a nozzle head located 100 m above the
Mylar.RTM. film. After solvent evaporation the stripes were
virtually colorless until wetted when they became faint green with
a basic aqueous sample for measurement.
EXAMPLE 8
[0099] A striped pH sensor constructed as in Example 7 was further
placed in the sampling device and measured with the Perkin Elmer
LS50-B in a manner similar to that described in example 6. In this
case, the excitation wavelength was set to 485 nm and emission
recorded at 530 nm while consecutive buffer samples corresponded to
pH 7.5, 7.1, 6.8, 7.1, and 7.5 were introduced to the sensor. The
reversible fluorescence quenching due to acidification of the
fluorescein sensor dye by the samples is as recorded in FIG.
10.
EXAMPLE 9
[0100] Using striping methods as described in examples 3,5 and 7, a
series of parallel sensor stripes for oxygen, carbon dioxide and pH
were laid down on a Mylar.RTM. film similar to that illustrated in
FIG. 1. A 150 m thick film of Mylar.RTM. with double sided adhesive
backing giving a total thickness of 210 m was punched with a series
of parallel cutouts transverse to the longitudinal direction of the
film to form intermediate web 26. This intermediate web was then
fixed to a clear film of Mylar.RTM. to form cover web 28, and a
series of holes punched, one at each end of the parallel cutouts.
In the final assembly step, the film containing the sensor stripes
was placed as the last sandwich layer on the bottom with the sensor
side in contact with the transverse cutouts on the intermediate
layer as shown in FIG. 4, thus forming sample chambers 118
approximately 210 m deep. For measurements and analyte
determinations, this sensor assembly was subsequently placed in an
instrument having several fiber optic splitter assemblies arranged
in parallel with the sample chambers. The appropriate color
excitation and collection optic was located directly below the
corresponding stripe to be measured as indicated in FIG. 5. As the
assembly containing the sensor stripes and sample chambers was
moved along, a bar containing an inlet and exit port was clamped
over the portal holes in the top clear Mylar.RTM. film (cover web
28) and an individual sample chamber was filled with a single
calibrant or sample. For demonstration purposes, ampuled vials of
the Certain.RTM. Plus standards by Chiron Diagnostics served as
both calibrants and samples with known values. These were opened
and aspirated into the sample wells over the sensor stripes. The
values for level 1 corresponded to pH 7.151, pCO2 68.9 mmhg and pO2
69.0 mmHg. The values for level 3 corresponded to pH 7.409, pCO2
40.1 mmHg and pO2 104.5 mmHg. The simultaneous response of the
sensors to a change in calibrant is illustrated in FIG. 11.
EXAMPLE 10
[0101] Using the sensor format and methodology described in example
9, a standard response curve was obtained for a single sensor
calibrated by three known Certain.RTM.
[0102] Plus standards corresponding to 71.6, 107.7 and 144.5 mm Hg
oxygen and is represented by the solid line shown in FIG. 12. The
optical sensor assembly was then advanced to a new position and
another different but known sample aspirated onto a fresh position
on each sensor stripe. These are represented by the single sensor
point responses. Table 1 shows a comparison of the measured values
calculated using the calibration algorithm. Although the
calibration was performed for one sensor, the algorithm was applied
to separate individual sensor positions along the stripe, each with
only a single measurement.
1 TABLE 1 Actual Level pO.sub.2 (mmHg) 71.6 107.7 144.5 Measured
Values with 73.4 113.9 142.6 Individual Sensors 74.3 110.6 133.1
101.8 156.3 Average 73.9 107.3 144.0
[0103] The foregoing description is intended primarily for purposes
of illustration. Although the invention has been shown and
described with respect to an exemplary embodiment thereof, it
should be understood by those skilled in the art that the foregoing
and various other changes, omissions, and additions in the form and
detail thereof may be made therein without departing from the
spirit and scope of the invention.
* * * * *