ACERAE AC AA
`(11] Patent Number:
`5,595,708
`United States Patent
`[45] Date of Patent:
`Jan. 21, 1997
`Berndt
`
`
`US005595708A
`
`119
`
`[56]
`
`Inventor: Klaus W. Berndt, Stewartstown, Pa.
`[75]
`[73] Assignee: Becton Dickinson and Company,
`Franklin Lakes, N.J.
`,
`
`436/52
`4/1994 Klinkhammer........
`5,304,492
`[54] SYSTEM FOR DETECTING BACTERIAL
`
`
`GROWTHIN A PLURALITY OF CULTURE 5,340,747=8/1994 Eden wees 436/172
`
`wae 235/462
`VIALS
`5,396,054
`3/1995 Kricheverctal.
`.....cccsssssessecnsee 422/52
`5,401,465
`3/1995 Smethers et al.
`FOREIGN PATENT DOCUMENTS
`0025350
`3/1981
`European Pat. Off. .
`0151855
`8/1985 European Pat. Off. .
`0523521
`1/1993 European Pat. Off. .
`Primary Examiner—James C. Housel
`[21] Appl. No.: 341,825
`Assistant Examiner—Rachel Heather Freed
`Attorney, Agent, or Firm—Alan W.Fiedler
`[22]
`Filed:
`Nov. 18, 1994
`[57]
`ABSTRACT
`Related U.S. Application Data
`‘System for detecting thepresenceof bacterial growth in a
`[62] Division of Ser. No. 113,444, Aug. 27, 1993, Pat, No.
`plurality of sample vials incorporates a single test stalion
`5,397,709.
`moveable along each of the plurality of sample vials. In one
`6
`embodiment, the sensor station is movably mounted on a
`Tint, (Ieccicessesessessecssssessseeses GOIN 21/01
`[51]
`
`[S52] US. Ch. oeceeesscssccssesessssesssseeuns 422/82.06; 422/82.05; rod, and that rod is movably mounted onapair of spaced
`436/164; 435/288.7, 356/337
`—_rods. The rod whichcarries thetest station may move along
`[58] Field of Search ou... 422/63, 67, 68.1,
`the spaced rods to change the location of the test station in
`422/82.057, 82.09; 436/47, 163, 164, 172;
`a first dimension andthetest station is moveable alongits
`356/337, 432, 436; 435/291, 240.2, 4
`rod to change location in a second dimension. In this way,
`:
`the test station may be moved through two dimensions to
`References Cited
`move serially to the location of each of the plurality of
`sample vials. In anotheraspectof this invention, a bar code
`U.S. PATENT DOCUMENTS
`is associated with each of the sample vials, and thetest
`3,773,426 11/1973 Mudd wcccscesstecssccsseesseessessessaees 356/205
`station makes a reading ofthat bar code concurrent with a
`5/1987 Deddenetal.
`4,665,036
`
`-- 435/301
`determination being made as to whetherthere is any bacte-
`4,772,453
`9/1988 Lisenbee......
`
`vee 422/52
`rial growth in the samplevial. In this way,it is ensured that
`al.
`.
`4,861,727
`8/1989 Hauenstein et
`eee
`the results of the evaluation of whether bacterial growth is
`
`3/1992 Leaback .....0..00.0...
`5,096,807
`
`~
`ongoing will be associated with the proper sample vial. In a
`3/1992 ‘Yokomori etal. ...
`5,096,835
`» 436/165
`:
`-
`:
`wae
`
`third aspect of this invention, the sample vial incorporates a
`11/1992 Di Guiseppietal.
`5,164,796
`w. 356/445
`:
`Sees
`:
`
`........00.
`5,168,766 12/1992 Stoffel
`_ 73/864.81
`plurality of distinct types of bacterial sensors. Thus,
`the
`
`5,234,665
`8/1993 Ohtaetal.
`...
`a 422/73
`advantages of each of several types of bacterial sensors may
`
`5,268,304 12/1993 Inman etal. .
`. 436/172
`be incorporated into a single sample vial.
`5,290,701
`3/1994 Wilkins........
`wee 435/312
`5,302,813
`4/1994 Goren ooeesseecesessesesseeeseecenees 235/462
`
`
`8 Claims, 13 Drawing Sheets
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`Agilent Exhibit 1239
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`Agilent Exhibit 1239
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`U.S. Patent
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`Jan. 21, 1997
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`Sheet 1 of 13
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`U.S. Patent
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`Jan. 21, 1997
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`5,595,708
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`Sheet 2 of 13
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`Agilent Exhibit 1239
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`Agilent Exhibit 1239
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`U.S. Patent
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`Jan. 21, 1997
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`Sheet 3 of 13
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`Agilent Exhibit 1239
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`Agilent Exhibit 1239
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`Agilent Exhibit 1239
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`U.S. Patent
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`Jan. 21, 1997
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`Sheet 5 of 13
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`Agilent Exhibit 1239
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`Jan. 21, 1997
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`Sheet 6 of 13
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`U.S. Patent
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`Jan. 21, 1997
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`Agilent Exhibit 1239
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`U.S. Patent
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`Jan. 21, 1997
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`Sheet 10 of 13
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`Jan. 21, 1997
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`This application in general relates to an improved system
`for monitoring a plurality of sample vials, and making a
`determination of whether the sample vials are experiencing
`bacterial growth.
`Sample vials are prepared by injecting a body fluid
`sample into a culture medium in a sample vial. The sample
`vial
`is then incubated, and tested for bacterial growth.
`Systems for detecting bacterial growth in sample vials are
`known, wherein a large numberof sample vials are repeat-
`edly and periodically tested for the presence of bacterial
`growth. Several types of sensors are known which have
`changing responses to a light input based on conditions
`within the sample vial. By monitoring the sensor response
`one can determine whether there is bacterial growth.
`Generally,
`in known sensors light is directed into the
`sample vial or sensor. Light reemerging from the sample
`vial, or from the sensor, is monitored to determine whether
`bacterial growth is occurring in the sample vial. Such
`sensors and associated methods of determination are known
`in the art, and the types of changes which indicate bacterial
`growth are known.
`In a preferred embodimentofthis invention, the bar code
`Knowntest systemstypically hold a large number of such
`is printed on a label associated with the sample vial, and a
`samplevials. In one example, they hold 240 sample vials.
`reference indicia is placed adjacentto the bar code. The bar
`With the known systems an individual
`light source, an
`code and referenceindicia are preferably printed on a single
`individual photodetector and the required wiring are asso-
`label. Thusthe reference indicia and bar code are alwaysat
`ciated with each sample vial. Thus, such systems are com-
`knownlocationsrelative to each other. Thetest station may
`plicated and expensive. Due to the large number oflight
`read the position of the reference indicia to ensure that the
`sourcesand detectors which are required, such systems have
`sensorstation is properly positioned relative to the bar code
`sometimesutilized less expensive light sources or detectors
`prior to reading the bar code. Further, the reference indicia
`than those which may be mostdesirable. Also, since several
`insures the test station is properly positioned relative to the
`hundredlight sources and photodetectorsare utilized within
`sample vial. In this way, the test station properly reads the
`each system,station to station variations are inevitable. That
`bar code, and eliminates misidcntification due to misposi-
`is, a light source associated withafirst station may emitlight
`tioning between thetest station and the bar codc.
`at a different intensity than the otherstations. Variation could
`In a most preferred embodimentofthis invention, the bar
`also occur between the photodetectors associated with the
`code is printed in a circular pattern, and the reference indicia
`hundredsofstations. This could result in potential variations
`is a circle concentric to the bar code. A circular bar code
`in readings between vials within the system. Suchvariations
`pattern provides the greatest length for bar code information
`are undesirable.
`:
`per unit space on the vial. Further, the circular bar code and
`reference indicia do not require any particular orientation of
`the sample vial about an axis of the sample vial relative to
`the test station.
`
`1
`SYSTEM FOR DETECTING BACTERIAL
`GROWTHIN A PLURALITY OF CULTURE
`VIALS
`
`This is a division of application Ser. No. 08/113,444,
`filed Aug. 27, 1993, now U.S. Pat. No. 5,397,709.
`
`BACKGROUND OF THE INVENTION
`
`Anotherproblem with thepriorart systemsis that the only
`identification of a vial located at a particular station within
`the system is by a manual bar code reading before the vial
`is placedinto the station. Thus, if an operator misplaces the
`vial within the station, there may be misidentification of the
`location of the vial within the station.
`types of
`Finally, as discussed above, there are several
`sensors which maybe utilized to determine the presence of
`bacterial growth. Each type of sensor has beneficial char-
`acteristics, and other characteristics that are undesirable.
`Further, certain types of bacteria are better detected by
`certain types of sensors. Thus, no one single type of sensor
`provides all desirable characteristics. Even so, the prior art
`has typically utilized vials with only a single type sensor
`incorporated into the vial.
`
`SUMMARYOF THE INVENTION
`
`2
`periodically and serially tests cach of the sample vials. In a
`preferred embodimentofthis invention,the samplevials are
`arranged in a two dimensional array. The test station is
`mounted on a frame which movesalong bothof the dimen-
`sions to test each vial.
`.
`
`Since a single test station tests each of the hundreds of
`vials, more expensive light sources and detectors may be
`incorporated while still reducing the cost of the overall
`system. More importantly, since only a single source and
`detector are utilized, station to station variations such as
`experienced with the prior art systems are eliminated.
`Several embodiments of this basic concept are disclosed
`in this application. The several embodiments incorporate the
`necessary apparatus for manyof the several types of sensors
`which may be utilized to test sample vials.
`In one preferred embodiment of this invention, the test
`station carries only an optical fiber. The optical fiber is
`operably connected to a source of light, and to a photode-
`tector. Thus, the moving test station frame need not carry
`any heavy equipment, or any cables to supply power to
`equipment mounted on the frame. Rather, the frame need
`only move the optical fiber.
`In a second aspectof this invention, a bar code is formed
`on the sample vial, and the test station includesstructure for
`reading the bar code from the sample vial. The bar code
`information is then positively associated with the test results
`from the vial. In this way, the results of the test are tied to
`the particular sample vial, and a misidentification of test
`results will not occur.
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`10
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`In a third aspect of this invention, a sensorpatchis placed
`on the bottom of the sample vial. The sensor patch includes
`a plurality of distinct types of sensors. As an example,
`sensors which respond to carbon dioxide concentration, pH
`level, oxygen level or other types of changes in the sample
`vial mayall be associated with each vial. In this way,a test
`station can test each of these various types of sensors, and
`make a more informed determination of whether the par-
`ticular sample vial is cxperiencing bacterial growth.
`These and other features of the present invention can be
`best understood from the following specification and draw-
`ings, of which the followingis a brief description.
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`55
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`60
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`In a disclosed embodimentof this invention, a single test
`station includes a light source and a light detector, and
`
`FIG. 1A showsa first embodimentof a system according
`to the present invention;
`
`Agilent Exhibit 1239
`Page 15 of 21
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`Agilent Exhibit 1239
`Page 15 of 21
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`FIG.1B is a partial end view, shown somewhat schemati-
`cally, of a system according to the present invention;
`FIG, 2 showsa vial bottom with a central bacterial sensor
`and a bar code pattern;
`FIG. 3 is a schematic of a control system associated with
`the system of FIG. 1;
`FIG. 4 shows a second embodimentof a system according
`to the present invention;
`FIG. 5 showsa third embodiment of a system according
`to the present invention;
`FIG.6 is a schematic of a control system associated with
`the system of FIG. 5;
`FIG. 7 showsa fourth embodimentof a system according
`to the present invention;
`FIG. 8 shows a vial bottom with a plurality of distinct
`sensors, and a bar code pattern;
`FIG.9 showsa vial bottom with a plurality of sensors, and
`a bar code pattern;
`FIG. 10 showsa fifth embodiment of a system according
`to the present invention;
`FIG. 11 depicts details of the rack area close to a vial
`bottom for the embodiment of FIG. 10;
`FIG.12 is a cross-sectional view taken along line 12-12
`of FIG. 11;
`FIG. 13 is a schematic illustrating the main optical and
`electronic componcnts of the embodiment of FIG. 10;
`FIG. 14 showsa sixth embodiment of a system according
`to the present invention;
`FIG. 15 is a planar view of a portion of the embodiment
`of FIG. 14;
`FIG. 16 is a cross-section of a light guide stub of the
`embodiment of FIG. 14; and
`FIG. 17 showsa vial bottom with a bar code pattern.
`
`DETAILED DESCRIPTION OF THE
`PREFERRED EMBODIMENTS
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`20
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`4
`arranged inside a known type incubator 44 used to promote
`microorganism growth.
`A single test station or carriage 43 is movedte test all of
`the vials. Light output is gencratcd from a single high energy
`light source, such as laser 46, and is serially directed at
`sensors 38 on a large numberof vials 32. Laser 46 and a
`detector module 64 are mountedontest station 43, which is
`movable as part of an XYtranslation stage 45. XY transla-
`tion stage 45 allows for movementof test station 43 along
`a rod 47 that is fixed to two guide blocks 49. The blocks 49
`move along perpendicularly arranged rods 51. If tipping
`racks 42, containing a total of 240 vials (12 rows and 20
`columns) are uscd, a single XY translation stage 45 must be
`able to address a maximum of 20 vials in one direction. As
`an alternative, two (or more) test stations can be used with
`each testing plural vials.
`In opcration, a system controller described below,dirccts
`test station 43 from a homeposition to a first vial station.
`Each of the vials 32 are then serially tested. As shown in
`FIG. 1B, the XYtranslation stage 45 movestest station 43,
`which carries laser 46. A plurality of sample vials 32 are
`arranged in a two dimensional array. Rods SI are positioned
`at the top and bottom ofthe vial array, and guide blocks 49
`carry rod 47 along rods 51. In one embodiment, rods 51
`could include a linear motor for driving guide blocks 49.
`Such linear motors are known, and may operate by passing
`current to move guide blocks 49 axially along bars 51.
`Similarly, test station 43 may move along bar 47 by use of
`a linear motor. In such a motor, test station 43 and guide
`blocks 49 would include magnets. By moving guide blocks
`49 alongrods 51, the locationoftest station 43 can be varied
`to the right and left as shown in FIG. 1B, and the up and
`downpositionof test station 43 as shown in FIG. 1B can also
`be varied by moving test station 43 along rod 47.
`Asstated above, rods 47 and 51 may incorporate linear
`motors which drive the guide blocks 49, andtest station 43
`and guide blocks 49 could incorporate magnets to be driven
`along the rods 47 and 51. Alternatively, any other known
`means of guiding the test station 43 through two dimen-
`sional movement maybeutilized. Knownx-y translators are
`used for navigation, plotting and printing. It is not the
`structure of the translator which is inventive, but rather the
`use of an x-y translator in this environment.
`As shown in FIG. 1A, a beam splitter 48 is mounted on
`test station 43 and splits an output beam 50 from laser 46
`into components 52 and 54. Reference beam component 52
`is directed toward a reference photodetector 55. Reference
`photodetector 55 measures the intensity of reference beam
`component 52 and generates a reference photocurrent value
`corresponding to the measured intensity. Output beam com-
`ponent 54 passes through mirror 56 having central aperture
`58,and is deflected off of a curved mirror 60 to contact and
`excite a sensor 38 of a selected vial 32. When excited by
`output beam component54, sensor 38 generates an emission
`which varies with the presence of biological activity in the
`illustrated embodiment, a fluorescence emission generated
`by the sensor increases in proportion to increased biological
`activity. Fluorescence intensity chemical sensors 38 are
`known which react to pH, oxygen concentration, carbon
`dioxide concentration, or in response to other biological
`activities.
`
`Agilent Exhibit 1239
`Page 16 of 21
`
`A first system 30 for intensity-based detection of micro-
`organisms is shown in FIG. 1A. System 30 holdsa plurality
`of vials 32, each sealed with a septum 34 and containing a
`mediuny/bodily fluid mixture 36. Typically, the body fluid is
`blood and the medium is prepared by known techniques.
`Each vial 32 contains an intensity-based chemical sensor 38
`disposed on an inner bottom surface 40. While an intensity
`sensor is used with this embodiment, other sensors which
`generate a selective emission or change their reflectivity,
`opacity, or color in the presence of biological activity are
`known and may also be used. In other systems, the sample
`may be scanned without the use of a separate sensor asso-
`ciated with the vial, e.g., scattered photon migration
`(“SPM”), as discussed further below.
`Two rows of vials 32 are arranged on each of several
`tipping racks. Tipping racks 42 are agitated to promote the
`growth of microorganisms within vials 32. Tipping racks 42
`may be biased or held in a known hold position, such asthat
`shownin FIG.1, while a determination of biological activity
`may be made. In someapplicationsit is desirable to hold the
`vials at an angle relative to the vertical to maximizethe level
`of fiuid. Any structure for moving the tipping rack, or
`holding it at the known position may be used. A plurality of
`An emission 62 fromaparticular sensor 38 is collected by
`tipping racks 42 are used since a tipping rack for as many as
`curved mirror 60, planar mirror 56 and directed toward an
`240 vials would have considerable mass. Racks 42 contain
`optical sensing detector module 64 where the emission is
`only vials 32 and no electronic components and, conse-
`monitored. Detector module 64 includes a spectral emission
`quently, no electrical wires. Vials 32 and tipping racks 42 are
`filter 66, and a high-sensitivity photodetector 68. Filter 66 is
`
`45
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`Agilent Exhibit 1239
`Page 16 of 21
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`5,595,708
`
`5
`used to block unwanted short-wavelength or. excitation
`radiationthat can affect readings. Photodetector 68 measures
`the intensity of emission 62 and generates a sensor photo-
`current value representative of the measured intensity.
`In one preferred embodiment, laser 46 is a green helium
`neon (HeNe) laser having a wavelength in the range of about
`543.5 nm, with approximately 1.5 mW ofoutput power. The
`diameter of output beam 50 should be no greater than about
`2 mm. The short-wavelength light and output power reacts
`well with a fluorescence sensor 38 in the presence of
`biological activity.
`With this embodiment, and all other embodiments dis-
`closed in this invention,the thrust of the invention is to the
`moving test station and associated structure, to bar code
`reading, and to a plural sensor embodiment as will be
`disclosed below. The parameters used to evaluate the read-
`ings madeonthe particular sensors are as known in theart.
`The present invention does not disclose any new testing
`logic. Rather, the detected emission, etc. are evaluated as
`known in the art to make a determination of whether a
`particular vial is experiencing bacterial growth.
`Thetest station 43 carried on XYtranslation stage 45 also
`includes a monitor photodiode 80 for measuringlight back-
`scattered from a bar code label 82 shown in FIG. 2,
`preferably placed on a bottom outer surface 84 ofvial 32. In
`the illustrated embodiment, bar code 82 includes a central
`opening 86 so that sensor 38 is exposed. In an alternative
`embodiment, bar code 82 may be printed on a label placed
`on bottom inner surface 40 which also includes sensor 38. A
`circular bar code pattern 88 comprises a plurality of marks
`89 extending radially outwardly from central opening 86.
`The marks 89 carry information with regard to thevial, the
`sample, or the patient associated with the sample. Bar code
`pattern 88 includes a reference indicia, here an outer con-
`centric circle 90. If used with a standard bloodculture vial,
`bar code pattern $8 can have a diameter of approximately 25
`mm. With such a diameter, the bar code information char-
`acterizing the vial can be distributed overa circle circum-
`ference length of 78.5 mm.Theeffective length ofa circular
`pattern is at a maximum comparedto otherpatterns for any
`given vial diameter.
`To read out the bar code information 88, small sinusoidal
`deflection signals are sent to XY translation stage 45 via X
`and Y output channels 106 and 108, as shown in FIG. 3. The
`two signals have equal amplitude, but show a phase differ-
`ence of 90 degrees. This results in a movement of output
`beam component 54 along the circumferenceofa circle. The
`amplitude of laser 46 is adjusted and output beam compo-
`nent 54 scans bar code pattern 88 including concentriccircle
`90. This light is backscattered to be read by photodiode80.
`If XY translation stage 43 is initially incorrectly positioned
`with respect to a vial 32, concentric circle 90 is used as a
`position encoder to more accurately position XY translation
`stage 45 with respectto a vial 32 before reading the bar code.
`The test station 43 moves until concentric circle 90 is read
`as being at an expected position relative to test station 43.
`The concentric circle 90 and bar code pattern 88 are printed
`onto a label at knownrelative positions.
`In the embodiment of FIG. 1A, output beam component
`54 is focussed onto vial bottom surface $4 using mirror 60.
`As mentioned above, the available geometrical length of the
`bar code pattern 88 is approximately 78.5 mm. Green HeNe
`lasers such as preferred for use in the FIG. 1 embodiment
`typically have beam diameters of 0.5 mm. With such dimen-
`sions, no extreme focussing is required to read out the bar
`code information. However, when necessary, stronger focus-
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`65
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`6
`sing is accomplished very easily by a simple lensattached to
`laser 46.
`
`Under some circumstances, a larger diameter for output
`beam component 54 may be desired. A small beam could
`result in sensor bleaching, since only a very small surface
`area of the sensor is exposed to the test light. To avoid such
`a result, output beam component 54 can be moved about a
`small circle. In other words, small sinusoidal deflection
`signals may besent to XYtranslation stage 45 via the X and
`Y channels 106 and 108 continuously. To move from the
`above described circular movements for reading the bar
`code, and then to switch from bar code reading to sensor
`reading, only the amplitude of the sinusoidal signals would
`have to be changed.
`Photodiode 80 is not necessary if bar code pattern 88 is
`printed using a dye which fluoresces within the same wave-
`length range as the emission from sensor 38. In this way, the
`same detector module 64, including photodetector 68, can be
`used for microorganism detection and for bar code reading.
`A major advantage of system 30 is that both laser 46 and
`detector module 64 may be moved closely to individual vials
`32. Therefore, an extremely high spatial resolution for the
`bar code reading and improvedlight detection sensitivity
`can be realized. Further, greater accuracy is achieved by
`using a single test station for many vials in place of
`individual devices for each vial. More expensive and precise
`instrumentation can be used at a reasonable cost. Further, the
`need for instrament calibration is greatly reduced, if not
`eliminated by the presentinvention.
`As known,vials 32 are continuously scanned until either
`there is a presenceof biological activity, or a predetermined
`period oftime,typically five days, has passed. Generally, the
`presence of biological activity in a vial is indicated by a
`pronounced change in the measured sensor emission 62.
`As shown schematically in FIG. 3, laser 46 generates an
`output beam 50. Beam splitter 48 splits output beam 50 into
`reference beam component 52 and output beam component
`54. With XYtranslation stage 45, shown in FIG.1, correctly
`positioned, output beam component 54 is directed to a
`preselected sensor 38 associated with a vial 32. The sensor
`then generates an emission 62. Photodetector 68 monitors
`emission 62 and generates a sensor photocurrent 92. Pho-
`tocurrent 92 is routed to a detector DC meter 94. An output
`96 from DC meter 94 is fed to a controller 98, such as a
`computer.
`Reference beam component 52 is directed to reference
`photodetector 55, which monitors reference beam compo-
`nent 52 and generates a reference photocurrent 100. Photo-
`current 100 is routed to areference DC meter 102. An output
`104 from meter 102 is also fed into controller 98. If the
`reference beam component 52 varies from an expected
`value, then the intensity of output beam 50 is also not as
`expected, or desired. The intensity of output beam 50 is
`adjusted as necessary.
`Controller 98 stores and analyzes outputs such as 96 and
`104 to make a determination concerning microorganism
`growth. As is known, controller 98 compares incoming data
`to earlier collected data. In addition to collecting and ana-
`lyzing information, controller 98 positions XY translation
`stage 45, with signals being sent through X output channel
`line 106 and Y output channel line 108. Knowncontrollogic
`is used for moving station 43 as desired. Thus, output beam
`component54 is directed serially from vial to vial allowing
`a determination of microorganism growth to be made for
`each vial 32. Backscattered light 99 may be detected by
`photodetector 80, as discussed above, to accurately position
`XY translation stage 45 and read the bar code information.
`
`Agilent Exhibit 1239
`Page 17 of 21
`
`Agilent Exhibit 1239
`Page 17 of 21
`
`

`

`: 5,595,708
`
`7
`
`Atypical green HeNelaser head of 0.2 mW output power
`only has a weight of 0.34 kg and a length of 25 cm.
`However, it still needs a high-voltage power cable. Similar
`considerations apply to high-sensitivity photodetectors such
`as photomultipliers. Thus,
`in the FIG. 1A embodiment
`cables must extend from the movingtest station to a fixed
`powersource and to the controller. These requirements are
`addressed by system 120 as illustrated in FIG. 4. A test
`station 122 is arranged on an XY translation stage 124. As
`in the cmbodiment of FIG. 1, XY translation stage 124
`allows for movementoftest station 122 along a rod 123 that
`is fixed to two guide blocks 125.In turn, the blocks 125 can
`move along perpendicularly arranged rods 127. Test station
`122 holds an output end 126 of an optical fiber 128, and an
`imaging lens 130. In this embodiment,laser 46 and detector
`module 64 are mounted to a pedestal 127 in close proximity
`to one another at a fixed position within system 120. The
`input end 132 of fiber 128 is mounted sothatit receives the
`full optical output of output beam component 54. Light
`travels along fiber 128 and reaches output end 126. Beam
`component 54 exiting fiber output end 126 is imaged onto
`bottom inner surface 40 of vial 32. An emission 62 reemerg-
`ing from surface 40 is re-focussed by lens 130 into output
`end 126, back along fiber 128 and out input end 132. The
`emission 62 that emerges from input end 132 is directed by
`mirrors 60 and 56 through filter 66 and to photodetector68.
`A major advantage of system 120 is that a minimum
`amount of mass has to be moved ontest station 122 for
`scanning an array of vials 32. In addition, no electrical
`cables or wires are required On movingteststation 122.
`Output end 126 may be located closely to the bottoms of
`vials 32. In so doing, an extremely high bar code resolution
`and goodlight detection sensitivity are achieved. Sensor and
`bar code reading may be performed by moving the test
`station 122 as described above with respect to the embodi-
`ment of FIG. 1A. As an alternative, a pair of fibers may be
`used, with one directing light at the sensor, and one receiv-
`ing cmissions from the sensor.
`A third system 140 according to the present invention for
`use with time decay detection of bacteria, is shown in FIG.
`5. System 140 is similar to system 30 illustrated in FIG. 1A.
`However, a different type of optical sensor, a fluorescence
`decay time sensor 142, is disposed on inner bottom surface
`40 of each of vials 32. Fluorescence decay time sensors are
`known which change their decay time in response to chang-
`ing pH, oxygen concentration, carbon dioxide concentration,
`or in response to other biological activities. Using this
`method,
`intensity measurements are replaced with time
`measurements, so intensity changes do not influence the
`results. For sensors 142 to work properly, a modulated light
`source 144 includes a high-frequency intensily modulator
`146 arranged between laser 46 and beam splitter 48. The
`laser may be the sameas that disclosed in the embodiment
`of FIG, 1A. Modulator 146 may be of any knowntype, such
`as acousto-optic, electro-optic or elasto-optic.
`Output 148 from modulated light source 144is split into
`components 150 and 152. Reference beam component 150
`is directed toward a reference photodetector 56 while output
`beam component 152 passes through planar mirror 56
`having a central aperture 58, and is deflected off of a curved
`mirror 60 to contact and excite a sensor 142 ofa selected vial
`32.
`
`A modulated emission 154 generated by a particular
`sensor 142 is time modulated in response to increasing
`biological activity. It is the modulation rather than intensity
`that is primarily monitored by detector module 64. As long
`as the modulation can be measured, a determination of
`
`20
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`35
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`40
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`45
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`55
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`60
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`8
`biological activity can be made. Therefore, minor vial mis-
`positioning, light source or detector module aging, and dark
`current changes such as those resulting from outside light
`leakage into incubator 44, are notcritical.
`Currently available fluorescence decay time sensors
`require high light modulation frequencies, typically above
`100 MH. In known systems with individual light sources at
`eachvial 32, green light emitting diodes (“LED”s)are used.
`LEDs cannot be modulated at such high frequencies. In
`apparatus 140, however, with laser 46 and modulator 146,
`high-frequency intensity modulation may be easily accom-
`plished. Since only a single laser is necessary, the usc of a
`laser which is more expensive then an LEDisstill practical.
`As shown schematically in FIG. 6, controller 98 controls
`modulator 146 using an amplifier 156. Controller 98 sends
`a signal 158 to amplifier 156, and an outputsignal 160 from
`amplifier 156 is sent to modulator 146. Beam splitter 48
`splits output beam 148 from modulator 146 into reference
`beam component 150 and output beam component 152.
`With XY translation stage 45 correctly positioned, output
`beam component 152is directed to a preselected sensor 142
`associated with a vial 32. Sensor 142 selectively generates
`a modulated sensor emission 154. Photodetector 68 moni-
`tors sensor emission 154 and generates a modulated photo-
`current 162 which is routed to a vector voltmeter 164.
`Reference photodetector 56 monitors reference beam com-
`ponent 150 and generates a modulated reference photocur-
`rent 166 whichis also routed to vector voltmeter 164. Vector
`voltmeter 164 compares photocurrents 162 and 166 to
`determine a sensor phase shift a modulation, and, optionally,
`sensor intensity. This information is fed into controller 98
`via voltmeter outputs 168 and 170 so that a determination
`may be reached regarding microorganism growth for each
`vial. As in the embodiment of FIG. 1A, besides storing
`inputs 168 and 170, computer 98 controls the positioning of
`XYtranslation stage 45 using X output channel line 106 and
`Y output channel line 108. Output beam