TOMMAND DUE TO THI WA TAMTHITTI US010036703B1

( 12 ) United States Patent Ng et al .

( 10 ) Patent No . : US 10 , 036 , 703 B1 ( 45 ) Date of Patent : Jul . 31 , 2018

( 54 ) PORTABLE LASER BIOSENSOR ( 56 ) References Cited U . S . PATENT DOCUMENTS ( 71 ) Applicant : The United States of America as

represented by the Secretary of the Navy , Washington , DC ( US )

( 72 ) Inventors : Kin Chiu Ng , Fresno , CA ( US ) ; Subrata Sanyal , Eastvale , CA ( US )

( 73 ) Assignee : The United States of America , as represented by the Secretary of the Navy , Washington , DC ( US )

6 , 091 , 502 A * 7 / 2000 Weigl . . . . . . . . . . . . . . . GOIN 27 / 44721 356 / 416

6 , 255 , 118 B17 / 2001 Alfano et al . 7 , 236 , 243 B26 / 2007 Beecroft et al . 7 , 286 , 222 B2 10 / 2007 Gardner , Jr . 7 , 636 , 154 B1 12 / 2009 LaValley et al . 7 , 851 , 251 B2 12 / 2010 Tseng et al . 8 , 502 , 168 B1 8 / 2013 Poteet et al . 8 , 548 , 174 B2 10 / 2013 Dufresne et al . 8 , 729 , 502 B1 5 / 2014 Klotzkin 8 , 849 , 380 B2 9 / 2014 Patwardhan 9 , 255 , 900 B2 2 / 2016 Fishbine et al .

2016 / 0084707 A1 3 / 2016 Scott et al . ( * ) Notice : Subject to any disclaimer , the term of this

patent is extended or adjusted under 35 U . S . C . 154 ( b ) by 0 days .

FOREIGN PATENT DOCUMENTS ( 21 ) Appl . No . : 15 / 712 , 120

WO WO 2015 / 061793 4 / 2015

OTHER PUBLICATIONS ( 22 ) Filed : Sep . 21 , 2017

Related U . S . Application Data ( 60 ) Provisional application No . 62 / 451 , 216 , filed on Jan .

27 , 2017 . Hossain et al . , “ Combined ' dual ' Absorption and Fluorescence Smartphone Spectrometers ” , Optics Letters , vol . 40 , No . 8 , Apr . 15 , 2015 , retrieved on Jun . 24 , 2016 from http : / / www . osapublishing . org / ol / abstract . cfm ? uri = 40 - 8 - 1737 . pdf .

( Continued )

Primary Examiner — Abdullahi Nur ( 74 ) Attorney , Agent , or Firm — Christopher A . Monsey

( 51 ) Int . CI . GOIN 21 / 00 ( 2006 . 01 ) GOIN 21 / 64 ( 2006 . 01 ) GOIN 21 / 31 ( 2006 . 01 ) GOIN 30 / 00 ( 2006 . 01 ) GOIN 21 / 65 ( 2006 . 01 )

( 52 ) U . S . CI . CPC . . . . . GOIN 21 / 6402 ( 2013 . 01 ) ; GOIN 21 / 3103

( 2013 . 01 ) ; GOIN 21 / 65 ( 2013 . 01 ) ; GOIN 2030 / 0095 ( 2013 . 01 )

( 58 ) Field of Classification Search CPC . . . . . GO1J 3 / 02 ; G01J 3 / 00 ; G01J 3 / 44 ; GOIN

21 / 65 ; GOIN 21 / 64 ; GO1N 21 / 645 ; GOIN 2021 / 64 ; GOIN 21 / 31

See application file for complete search history .

( 57 ) ABSTRACT A hand - held laser based biosensor including modular com ponents , combining absorption spectrophotometry and molecular fluorescence spectrophotometry , and including a sample cell module having an improved absorption path length .

46 Claims , 8 Drawing Sheets

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( 56 ) References Cited

OTHER PUBLICATIONS Baird et al . , “ Compact , Self - Contained Optical Spectrometer ” , Applied Spectroscopy , vol . 49 , No . 11 , 1995 , pp . 1699 - 1704 , retrieved on Jun . 24 , 2016 from http : / / www . osapublishing . org / as / abstract . cfm ? uri = as - 49 - 11 - 1699 . pdf . Ocean Optics , “ Maya2000Pro Data Sheet for Versions 3 . 00 . 1 and Above ” , retrieved on Jun . 23 , 2016 from http : / / oceanoptics . com / wp - content / uploads / OEM - Data - Sheet - Maya2009Prov3 . pdf . Horiba Scientific , “ Fluorolog - 3 , How to build a Spectrofluorometer ” , retrieved on Jun . 24 , 2016 from http : / / horiba . com / fileadmin / uploads / Scientific / Documents / Fluorescence / flogcat . pdf . Soper et al . , " Molecular Fluorescence , Phosphorescence , and Chemiluminescence Spectrometry " , Analytical Chemistry , vol . 66 , No . 12 , Jun . 15 , 1994 , retrieved on Jun . 23 , 2016 from http : / / pubs . acs . org / dei / abs / 10 . 1021 / ac00084a016 ? journalCode + ancham & . pdf . Strianese et al . , “ Absorption into Fluoresence . A method to sense biologically relevant gas molecules ” , Nanoscale , 2011 , 3 , 298 - 302 ,

retrieved on Jun . 23 , 2016 from http : / / www . ncbi . nlm . nih . gov / pubmed / 21060958 . pdf . Clark et al . , “ Biological Detection System Technologies — Technol ogy and Industrial Base Study ” , Feb . 2001 , retrieved on Jun . 23 , 2016 from http : / / www . acq . osd . mil / mibp / natibo / docs / BioDetectReport - 2 . pdf . Thrush et al . , “ Integrated bi - fluorescence sensor ” , Journal of Chro matography 1013 ( 2003 ) 103 - 110 , retrieved on Jun . 23 , 2016 from http : / / www . breault . com / sites / default / files / knowledge _ base / wp _ els _ 001 _ integrated _ bio - fluorescence . pdf . So et al . , “ Fluorescence Spectrophotometry " , Encyclopedia of Life Sciences , 2002 , retrieved on Jun . 23 , 2016 from http : / / web . mit . edu / solab / Documents / Assets / So - Fluorescence % 20pectrophotometry . pdf . Cetin et al . , “ Handheld high - throughput plasmonic biosensor using computational on - chip imaging ” , Light : Science & Applications , 2014 , retrieved on Jun . 23 , 2016 from http : / / www . nature . com / lsa / journal / v3 / nl / pdf / lsa20143a . pdf .

* cited by examiner

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US 10 , 036 , 703 B1

PORTABLE LASER BIOSENSOR housing and configured to generate excitation energy , illus tratively a laser beam . The hand - held biosensor further

CROSS - REFERENCE TO RELATED includes a sample cell module including a sample cell APPLICATION module housing defining a chamber configured to receive a

5 solution , a sample inlet in fluid communication with the The present application claims priority to U . S . Provisional chamber , and a waste outlet in fluid communication with the

Patent Application Ser . No . 62 / 451 , 216 , filed Jan . 27 , 2017 , chamber . A first releasable coupling is positioned between the disclosure of which is expressly incorporated herein by the radiation emitting module and the sample cell module . reference . An absorption detector is configured to detect emission

10 intensity produced by the electromagnetic radiation passing STATEMENT REGARDING FEDERALLY through the solution in the sample cell chamber and produce

SPONSORED RESEARCH OR DEVELOPMENT absorption spectral data in response thereto , and produce absorption spectral data in response thereto , the absorption

The invention described herein includes contributions by detector including an absorption detector spectral filter . A one or more employees of the Department of the Navy made 15 fluorescence detector is configured to detect molecular emis in performance of official duties and may be manufactured , sions produced by the laser excited molecules as the laser used , and licensed by or for the United States Government beam passes through the solution in the sample cell chamber without payment of any royalties thereon . This invention and produces fluorescence spectral data in response thereto , ( Navy Case 200 , 408 ) is assigned to the United States the fluorescence detector including a fluorescence detector Government and is available for licensing for commercial 20 spectral filter . A processor is in electrical communication purposes . Licensing and technical inquiries may be directed with the absorption detector and the fluorescence detector , to the Technology Transfer Office , Naval Surface Warfare the processor being configured to receive the absorption Center Corona Division , email : CRNA _ CTO @ navy . mil . spectral data from the absorption detector and the fluores

cence spectral data from the fluorescence detector . A display BACKGROUND AND SUMMARY OF THE 25 module is in electrical communication with the processor ,

DISCLOSURE the display module including a display module housing , an absorption display supported by the display module housing

The present disclosure relates generally to methods and and configured to provide an indication of the absorption apparatuses for detecting and identifying substances within spectral data from the absorption detector , and a fluores a sample fluid . More particularly , the present invention 30 cence display supported by the display module housing and relates to a portable , illustratively pocket wearable , laser configured to provide an indication of a fluorescence spec based biosensor including modular components , and com tral data from the fluorescence detector . A power source is in bining molecular absorption spectrophotometry and molecu communication with the processor . A second releasable lar fluorescence spectrophotometry . coupling is positioned between the display module and the Molecular absorption spectrophotometry ( MAS ) and 35 sample cell module .

molecular fluorescence spectrophotometry ( MFS ) are well According to another illustrative embodiment of the pres established analytical techniques . More particularly , a vari ent disclosure , a hand - held biosensor includes an outer ety of dedicated molecular absorption spectrophotometers casing extending between a proximal end and a distal end , and dedicated molecular fluorescence spectrophotometers and a radiation emitting module supported proximate the are available from a variety of chemical instrument manu - 40 proximal end of the outer casing . A display module is facturers . Some of these devices are portable . While simul - supported proximate the distal end of the outer casing , and taneous operation of MAS and MFS has been performed in the sample cell module is supported intermediate the radia research laboratories for solving specialized , fundamental tion emitting module and the display module . The sample problems ; there are no known portable combination absorp - cell module includes a sample cell module housing defining tion - fluorescence spectrophotometers available in the mar - 45 a chamber and including a light entry port , a light exit port ket for the field testing of substances within a fluid sample . axially spaced by the chamber from the light entry port , a

There remains a need for the efficient field testing of longitudinal axis defined by the light entry port and the light substances within a fluid sample , combining both molecular exit port , and a reflective surface configured to reflect light absorption spectrophotometry ( MAS ) and molecular fluo - from the radiation emitting module transverse to the longi rescence spectrophotometry ( MFS ) , in a single spectropho - 50 tudinal axis . The hand - held biosensor further includes an tometer , for performing both absorption and fluorescence absorption detector , a fluorescence detector , and a processor measurements and providing rapid results to the user . in electrical communication with the absorption detector and

The present invention relates to a hand - held laser based the fluorescence detector , the processor being configured to biosensor that illustratively is portable ( e . g . , pocket wear - receive absorption spectral data from the absorption detector able by a user ) , is modular ( e . g . , interchangeable modules ) , 55 and fluorescence spectral data from the fluorescence detec has improved selectivity ( e . g . , combined molecular absorp - tor . The sample cell module includes an axial length between tion spectrophotometry and molecular fluorescence spectro - the light entry port and the light exit port , the axial length photometry ) , and has improved sensitivity ( e . g . , long b eing at least about 5 centimeters . absorption path length and long emission ( fluorescence ) According to another illustrative embodiment of the pres region ) . Such a biosensor may find use in a variety of 60 ent disclosure , a method of constructing a hand - held bio applications including , for example , in the military , food and sensor includes the steps of providing a sample cell module drug industries , and with first responders ( e . g . , police and including a sample cell module housing having a side wall firefighters ) . defining a chamber and extending longitudinally between a

According to an illustrative embodiment of the present proximal end and a distal end , a sample inlet in communi disclosure , a hand - held biosensor includes a radiation emit - 65 cation with the chamber , and a waste outlet in communica ting module including light emitting module housing , and a tion with the chamber . The method further includes the steps light source received within the radiation emitting module of providing a solution within the chamber of the sample cell

US 10 , 036 , 703 B1

module , providing a radiation emitting module including a BRIEF DESCRIPTION OF THE DRAWINGS radiation emitting module housing extending longitudinally between the proximal end and the distal end , a light source A detailed description of the drawings particularly refers received within the radiation emitting module housing and to the accompanying figures , in which : configured to generate an excitation laser beam , and releas - 5 FIG . 1 is a perspective view of an illustrative portable , ably coupling the distal end of the radiation emitting module illustratively pocket wearable , biosensor of the present dis housing to the proximal end of the sample cell module housing . The method further includes the steps of providing FIG . 2 is a longitudinal cross - sectional view of the an absorption detector at the distal end of the sample cell , the biosensor of FIG . 1 ; absorption detector configured to detect the intensity of the 10 FIG . 3 is an exploded longitudinal cross - sectional view of laser beam ( energy ) passing through the solution in the the biosensor of FIG . 2 ; sample cell chamber , the absorption detector including an absorption detector spectral filter , and providing a fluores FIG . 4 is a cross - sectional view similar to FIG . 2 , showing cence detector configured to detect emissions produced by the light source emitting an excitation energy , illustratively laser excited molecules as the laser beam passes through the 15 a laser beam passing through a sample chamber 10 an rough the 15 a laser beam passing through a sample chamber to an solution of the sample cell chamber , the fluorescence detec absorption detector and a reflective surface reflecting fluo tor including a fluorescence detector spectral filter . The rescence to a fluorescence detector ; method also includes the steps of providing a processor in FIG . 5 is a block diagram illustrating interaction between electrical communication with the absorption detector and various components of the biosensor of FIG . 1 ; and the fluorescence detector , the processor configured to 20 FIGS . 6A - 6C are perspective views illustrating various receive absorption spectral data from the absorption detector steps of operation of the biosensor of FIG . 1 . and fluorescence spectral data from the fluorescence detec tor . The method further includes the steps of providing a DETAILED DESCRIPTION OF THE DRAWINGS display module in electrical communication with the pro cessor , the display module including the display module 25 Various embodiments of the invention described herein housing extending between a proximal end and a distal end , are not intended to be exhaustive or to limit the invention to an absorption display supported by a display module hous - precise forms disclosed . Rather , the embodiments selected ing and configured to provide an indication of the absorption for description have been chosen to enable one skilled in the spectral data , and a fluorescence display supported by the art to practice the invention . display module housing and configured to provide an indi - 30 Referring initially to FIGS . 1 - 4 , an illustrative hand - held cation of the fluorescence spectral data . The method further biosensor 10 is shown for use in detecting substances ( e . g . , includes the steps of releasably coupling the distal end of the biological molecules , often referred to as biomolecules or sample cell module housing to the proximal end of the bio - agents ) within a sample fluid ( e . g . , gas or liquid ) . display module housing , and providing a power source in Illustratively , the biosensor 10 includes an outer casing 12 electrical communication with the processor . 35 extending along a longitudinal axis 14 between a proximal

According to further illustrative embodiment of the pres - ( or lower ) end 16 and a distal ( or upper ) end 18 . The ent disclosure , a method of detecting a substance within a biosensor 10 is configured to have an ergonomic size , shape fluid sample using a hand - held biosensor includes the steps and weight , thereby facilitating operation by a user . With of providing a sample cell module including a sample cell reference to FIG . 2 , the outer casing 12 is configured to have module housing having a side wall defining a chamber and 40 dimensions substantially like that of a conventional laser extending along a longitudinal axis between a proximal end pointer and , as such , is small enough to be used or operated and a distal end , a sample inlet in fluid communication with while being held in the hand or hands of a user , and may be the chamber , and a waste outlet in fluid communication with stored within a conventional pocket of the user . More the chamber , and fluidly coupling a waste collection vial to particularly , the outer casing 12 is illustratively cylindrical the waste outlet . The method further includes the steps of 45 and has an axial length ( 1 ) of less than about 20 centimeters injecting a sample solution through the sample inlet into the ( 7 . 87 inches ) and an outer diameter ( d ) of less than about 4 chamber of the sample cell module , and generating an centimeters ( 1 . 57 inches ) . In a further illustrative embodi electromagnetic radiation with a laser light source . The ment , the outer casing 12 has an axial length ( 1 ) of less than method also includes the steps of directing the electromag about 16 centimeters ( 6 . 30 inches ) and an outer diameter ( d ) netic radiation through the sample solution in the chamber of 50 of less than about 3 centimeters ( 1 . 18 inches ) . In one the sample cell , absorbing the electromagnetic radiation by illustrative embodiment , the outer casing 12 has an axial the sample solution , sensing the intensity of the electromag - length ( 1 ) of approximately 14 . 2 centimeters ( 5 . 6 inches ) , netic radiation absorbed by the sample solution by an and an outer diameter ( d ) of approximately 2 centimeters absorption detector , and generating an absorption signal of ( 0 . 79 inches ) . A representative weight of the hand - held interest from the absorption detector . The method further 55 biosensor 10 is approximately 4 ounces . An external clip 20 includes the steps of producing fluorescence emissions from is illustratively coupled proximate the distal end 18 of the the sample solution , sensing the fluorescence emissions by outer casing 12 for coupling to clothing of a user , for a fluorescence detector , and generating a fluorescence signal example to a shirt pocket ( not shown ) . of interest from the fluorescence detector . The method also The biosensor 10 illustratively includes a plurality of includes steps of displaying an indication of the absorption 60 releasably coupled component sections or modules , includ signal of the absorption display , and displaying an indication ing a radiation emitting module 22 , a sample cell module 24 , of the fluorescence signal on the fluorescence display . an absorption detector module 26 , and a display module 28 ,

Additional features and advantages of the present inven - all aligned along the longitudinal axis 14 . Illustratively , the tion will become apparent to those skilled in the art upon radiation emitting module 22 is positioned adjacent the consideration of the following detailed description of the 65 proximal end 16 of the biosensor 10 , and the display module illustrative embodiments exemplifying the best modes of 28 is positioned adjacent the distal end 18 of the biosensor carrying out the invention as presently perceived . 10 .

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With reference to FIGS . 3 and 4 , the radiation emitting BA5367U Lithium Sulfur Dioxide ( 1 inchx1 inchx0 . 075 module 22 illustratively includes a radiation emitting mod - inch ) battery produced by Saft SA in France may be used . ule housing 30 having a cylindrical side wall 32 extending Another illustrative battery is the BA5367U battery together between a proximal end 31 and a distal end 33 . An end cap with a DC / DC High Voltage DBC - Series ( 0 . 83 inchx0 . 83 34 is illustratively threadably coupled to the proximal end 31 5 inchx0 . 32 inch ) Module , available from Laser Components of the side wall 32 . Both the side wall 32 and the end cap 34 USA , for raising the voltage output . are illustratively formed of a durable , light weight material , first or proximal power switch 52 is illustratively in such as anodized aluminum or thermoplastic . electrical communication with the proximal battery 51 and

The radiation emitting module 22 is configured to emit is supported by the end cap 34 proximate the proximal end electromagnetic radiation ( e . g . , laser light ) toward the 10 16 of the outer casing 12 . The proximal power switch 52 sample cell module 24 . Illustratively , a radiation source , may be a toggle push button configured to be depressed by such as a laser diode 36 and a laser diode driver 37 are a user . positioned within the radiation emitting module housing 30 The sample cell module 24 is illustratively positioned and is configured to emit electromagnetic radiation . More distal of the radiation emitting module 22 and includes a particularly , the laser diode 36 is configured to emit an 15 sample cell module housing 54 defining the sample cell ultraviolet radiation laser beam 38 in an axial direction ( i . e . , chamber 40 . A sample inlet 58 is positioned at a proximal along the longitudinal axis 14 as shown in FIG . 4 ) toward a end 60 of the housing 54 and is in selective fluid commu sample cell chamber 40 defined by the sample cell module nication with the sample cell chamber 40 . Similarly , a waste 24 . As further detailed herein , in an illustrative universal outlet 62 is positioned at a distal end 64 of the housing 54 embodiment of biosensor 10 , the wavelength of the laser 20 and is in selective fluid communication with the sample cell beam 38 emitted by the laser diode 36 is illustratively chamber 40 . Illustratively , both the sample inlet 58 and the between 260 nm and 290 nm , and more particularly about waste outlet 62 may include a resilient septum 66 and 68 , 275 nm , for native absorption and native fluorescence . In an respectively . As further detailed herein , the septum 66 of the illustrative selective embodiment of biosensor 10 , the wave - sample inlet 58 is configured to receive the tip 70 of a length of the laser beam 38 emitted by the laser diode 36 will 25 conventional syringe 72 including a plunger 74 slidably be centered approximately at the excitation wavelength for received within a barrel 76 . Similarly , the septum 68 of the absorption and fluorescence of the labeled biomolecule . The waste outlet 62 may receive the tip 78 of tubing 80 which is laser diode 36 is illustratively soldered to the laser diode coupled to a collection device , such as a vial 82 ( FIG . 6A ) . driver 37 . Illustratively , a suitable laser diode driver 37 is the The sample cell module housing 54 illustratively includes CW Laser Diode Driver LSC - 025 ( 0 . 82 inchesx0 . 29 inchesx 30 an outer cylindrical side wall 84 extending between the 0 . 20 inches ) , available from Laser Components USA , Inc . of proximal end 60 and the distal end 64 . A first or proximal Bedford , N . H . end wall 90 is coupled to the proximal end 60 of the side

A lens 41 is illustratively positioned proximate to , and wall 84 , and a second or distal end wall 92 is coupled to the distal of , the laser diode 36 . More particularly , the lens 41 is distal end 64 of the side wall 84 . The side wall 84 and the positioned between the laser diode 36 and the sample cell 35 end walls 90 and 92 are illustratively formed of a durable , chamber 40 . The illustrative laser diode driver 37 is operably light weight material , such as anodized aluminum or ther coupled to and positioned proximal of the laser diode 36 . moplastic . The inner surface 100 of the sample cell chamber The lens 41 may be of conventional design ( e . g . , formed of 40 is illustratively darkened , illustratively by application of a quartz material ) and is used for collimating the laser beam a dark coating and / or texturing . More particularly , the inner 38 to within a desired diameter , illustratively less than about 40 surface 100 may be defined by the sidewall 84 and the end 3 millimeters . walls 90 and 92 , and illustratively darkened for avoiding

A spatial filter 42 is illustratively positioned distal of the reflective and scattered light . lens 41 , between the lens 41 and the sample cell chamber 40 . light entry port 96 is illustratively formed within the The spatial filter 42 may be of conventional design for proximal end wall 90 , while a light exit port 98 is illustra blocking scattered light and thereby removing spatial noise . 45 tively formed within the distal end wall 92 . Illustratively , the More particularly , the illustrative spatial filter 42 includes a light entry port 96 and the light exit port 98 are aligned along mask 44 including a center opening 46 ( illustratively having the longitudinal axis 14 , and each has an illustrative diam a diameter no greater than 3 millimeters ) to allow only the eter of about 5 millimeters centimeters ( 0 . 20 inches ) . The laser beam 38 to pass therethrough , and blocking scattered sample cell module housing 54 is illustratively sealed from laser light . 50 external light , except for the light entry port 96 and the light

A first spectral filter 48 is illustratively positioned distal of exit port 98 . the spatial filter 42 . More particularly , the spectral filter 48 The sample cell module housing 54 illustratively defines is illustratively positioned between the spatial filter 42 and a quartz sample cell chamber 40 . An inner surface 100 of the the sample cell chamber 40 . The spectral filter 48 may sample cell module housing 54 may be textured for self comprise a band - pass / interference filter that permits only 55 cleaning . In an illustrative embodiment , the inside surface radiation from the laser diode 36 to pass therethrough . For 100 of the sample cell module housing 54 is nano - textured example , the spectral filter 48 allows only radiation ( i . e . , using , for example , ultra - short pulsed laser ( USPL ) beams or laser beam 38 ) of the laser diode 36 to pass therethrough other known techniques to render the inner surface super ( e . g . , having a wavelength of about 275 nm + / – 15 nm in the hydrophobic ( i . e . , water repelling ) and / or super - lipophobic illustrative universal embodiment of biosensor 10 ) . 60 ( i . e . , repelling organic solvents / fats ) . The quartz sample cell

The radiation emitting module 22 further includes a first chamber 40 , as opposed to plastic vials or cells , reduces the or proximal power source 50 illustratively in electrical chances of reaction and degradation , and promotes resulting communication with the radiation emitting module 22 . The prolonged usage and duration when used with various first power source 50 may comprise a proximal battery 51 bio - agents . The walls of the quartz sample cell chamber 40 releasably supported within the end cap 34 . The proximal 65 allows the laser light ( illustratively 275 nm wavelength ) and battery 51 is illustratively a rechargeable button cell battery . the fluorescence radiation ( illustratively 345 nm wave In one illustrative embodiment , a military grade 3 . 0 Volt Saft length ) to pass through .

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With further reference to FIGS . 3 and 4 , a fluorescence The absorption detector module 26 illustratively includes detector 102 is illustratively supported adjacent a first side an absorption detector module housing 120 . The absorption 104 of the sample cell module housing 54 . More particularly , detector module housing 120 illustratively includes a cylin the fluorescence detector 102 may comprise a strip detector drical side wall 122 extending between a proximal end 124 coupled to the side wall 84 of the sample cell module 5 and a distal end 126 . The side wall 122 is illustratively housing 54 . formed of a durable , light weight material , such as anodized

Illustratively , the fluorescence detector 102 is configured aluminum or thermoplastic . to detect electromagnetic radiation having a wavelength of An absorption detector 128 is received within the absorp interest , illustratively between 200 nm and 1000 nm . In one tion detector module housing 120 , and is configured to illustrative embodiment , the fluorescence detector 102 is 10 detect emission intensity / energy produced by electromag configured to detect electromagnetic radiation having a netic radiation passing through a solution in the sample cell wavelength of about 345 nm . In one illustrative embodi - chamber 40 . In response , the detector 128 produces absorp ment , the fluorescence detector 102 is a high quantum - tion spectral data . In an illustrative embodiment , the absorp efficiency avalanche photodiode - array , such as 1 . 5 mm tion detector 128 is an absorption spot detector . As further UV - VIS ( 200 - 1000 nm ) , Si APD , Stock No . 59 - 184 , avail - 15 detailed herein , the absorption detector 128 in a universal able from Edmund Optics Inc . of Barrington , N . J . embodiment of the biosensor 10 may comprise a deep

A fluorescence filter 106 is illustratively supported inter - ultraviolet ( DUV ) photodiode . Since the absorption detector nally of the fluorescence detector 102 . More particularly , the 128 directly senses the laser beam , sensitivity is not an issue . fluorescence filter 106 is illustratively positioned between In a selective embodiment of the biosensor 10 that employs the sample cell chamber 40 and the fluorescence detector 20 a longer wavelength , a wide variety of known diodes are 102 . Illustratively , the fluorescence filter 106 comprises a readily available . spectral filter , such as a narrow band - pass filter configured to Illustratively , the absorption detector 128 is configured to allow only fluorescence light of a wavelength of interest to detect electromagnetic radiation having a wavelength of pass therethrough . In an illustrative universal embodiment between about 200 nm and 980 nm , and more particularly a of the biosensor 10 , the fluorescence filter 106 allows only 25 wavelength of about 275 nm . More particularly , the absorp radiation having a wavelength between about 295 nm and tion detector 128 may comprise a silicon photodiode , such 420 nm , and more particularly about 345 nm , to pass as 5 . 7 mm ? DUV Photodiode with Ceramic Housing , Stock therethrough . No . 84 - 982 , available from Edmund Optics Inc . of Bar

A curved reflective surface 108 is illustratively supported rington , N . J . adjacent a second side 110 of the sample cell module 30 second spectral filter 130 is supported internally or housing 54 , opposite the fluorescence detector 102 adjacent proximal of the absorption detector 128 . More particularly , the first side 104 of the sample cell module housing 54 . the second spectral filter 130 is positioned between the Illustratively , the curved reflective surface 108 is defined by sample cell chamber 40 and the absorption detector 128 . The a polished surface or a mirror , illustratively a concave or spectral filter 130 may comprise a band - pass / interference spherical mirror 112 configured to curve - fit with the sample 35 filter that permits only radiation from the laser diode 36 to cell module housing 54 . The mirror 112 is configured to pass therethrough . For example , the spectral filter 130 collect and focus light or fluorescence emission 113 ( FIG . 4 ) allows only radiation of the laser diode 36 ( e . g . , about 275 toward the fluorescence detector 102 . The reflective surface nm + / - 15 nm in the illustrative universal embodiment of 108 is illustratively configured to collect and focus light 113 biosensor 10 ) . with a wavelength of about 345 nm . 40 More particularly , the illustrative absorption detector 128

In the illustrative embodiment universal biosensor 10 , the senses the intensity of the laser beam 38 . In operation , the laser beam 38 is an electromagnetic radiation of approxi - original intensity of the laser beam 38 ( for example , 275 nm mately 275 nm wavelength , which is used to uniquely excite wavelength ) passes through a blank solution in the sample amino - acids tryptophan ( Trp ) and tyrosine ( Tyr ) in the cell chamber 40 , is detected from the absorption detector sample solution in sample cell chamber 40 . Fluorescence 45 128 , is registered and showed as zero absorption signal at an emission 113 ( the 345 nm wavelength of light emitted from absorption display 152 . An absorption signal higher than excited amino - acids tryptophan and tyrosine in this z ero represents a partial absorption of the laser beam 38 . The example ) is collected with mirror 112 and reflected to the approximately 275 nm wavelength in this example indicates direction of the fluorescence detector 102 . A blank solution the presence of the amino acids tryptophan ( Trp ) and / or within the sample cell chamber 40 generates no fluorescence 50 tyrosine ( Tyr ) in the sample solution in the sample cell detection , provides background from the fluorescence detec - chamber 40 , and therefore , the presence of biomolecules . tor 102 , and gives a background - subtracted zero fluores . The sample cell module 24 facilitates high sensitivity by cence reading at a fluorescence display 150 . Any fluores - providing a long absorption path length and a long emission cence signals above zero represents detection at the ( fluorescence ) region . Absorption spectrophotometry is fluorescence detector 102 of the 345 nm wavelength of light , 55 defined by Beer ' s Law according to the equation : A = a : b . c , characteristically representing the presence of amino - acids where A is the absorbance signal ( the remaining intensity of tryptophan and / or tyrosine in the sample solution within the the source radiation detected ) , a is the unique spectral sample cell chamber 40 and symbolizing detection of absorption of monochromatic source radiation ( illustratively biomolecules . 275 nm laser light ) for the example biomolecule at that

A first releasable coupling 114 is positioned between the 60 wavelength , b is the absorption cell radiation path length , radiation emitting module 22 and the sample cell module 24 . and c is the concentration of the sample solution . Illustratively , internal threads 116 on the distal end 33 of the As may be appreciated by Beer ' s Law , the absorbance radiation emitting module 22 engage external threads 118 on signal ( A ) increases when the absorption cell radiation cell the proximal end 60 of the sample cell module 24 . Other path length ( b ) increases . With reference to FIG . 4 , the path releasable couplings may be substituted for cooperating 65 length ( b ) of the sample cell in the illustrative biosensor is threads , including , for example bayonet couplings , resilient at least about 5 centimeters ( 1 . 96 inches ) . This path length fingers , lock washers , etc . provides improved signal strength of about five times over

US 10 , 036 , 703 B1 10

known biosensors having reduced path lengths ( approxi solution is dye - labeled prior to injection into the sample cell mately 1 centimeter ( 0 . 39 inches ) ) . chamber 40 . Certain organic - dyes or semi - conductor quan

In fluorescence spectrophotometry , a molecule absorbs its tum dots preferentially bind to particular biomolecules / unique excitation radiation and emits a red shifted charac - proteins . The dye - labeled or quantum dot - labeled protein teristic radiation . The intensity of this emission is monitored 5 has a characteristic absorption ( excitation ) and fluorescence by the fluorescence detector 102 . Illustratively , a biomol ( emission ) wavelength profile . ecule absorbs laser light 38 of wavelength 275 nm and emit In one illustrative embodiment of the biosensor 10 con fluorescence light 113 of wavelength 345 nm . figured as a selective sensor , the laser diode 36 is configured Typically , with a fluorescence spectrophotometer , the to emit electromagnetic radiation having a wavelength of emission from a sample cell having a volume less than 1 cm3 10 between about 400 nm and about 410 nm . The fluorescence ( 0 . 06 in ) generated with diffused - lamp - excitation , is col measurement is 465 nm as defined by the fluorescence lected right - angle to the source - radiation and focused onto a band - pass filter 106 . The proper combination of laser diode spot - detector . The biosensor 10 of the present disclosure employs an intense , directional , laser - beam for optically 36 and filter 106 depends on the detection of the thin excitation of a relatively long ( approximately 5 cm 155 biomolecule ( s ) of interest . Modules or component sections ( 1 . 96 inches ) ) sample path ; and generates a long emission are illustratively constructed for a particular application , for image that is collected at a right angle to the laser source by example , optimal wavelengths for excitation , and for the concave reflective mirror ( surface ) 112 and focused onto absorption and fluorescence ( detection ) measurements . the fluorescence strip detector 102 . The detector and the A second releasable coupling 140 is positioned between emission image has an approximately overlapped dimen - 20 the sample cell module 24 and the absorption detector sion , resulting in high sensitivity . module 26 . Illustratively , internal threads 142 on the distal As further detailed herein , the biosensor 10 may be used end 64 of the sample cell module 24 engage external threads

as a universal sensor for detecting all biomolecules , and / or 144 on the proximal end 124 of the absorption detector a selective sensor for detecting only particular biomolecules . module 26 . Other releasable couplings may be substituted All biomolecules and biological cells ( e . g . , spores , human 25 for cooperating threads , including , for example bayonet and bacterial cells , etc . ) have proteins with their constituents couplings , resilient fingers , lock washers , etc . being amino acids . Amino acids bond together to form controller 145 , including a processor 146 and a memory proteins . The amino acids phenylalanine ( Phe ) , tryptophan 147 , is illustratively in electrical communication with the ( Trp ) and tyrosine ( Tyr ) can be monitored for " native ” ( i . e . , fluorescence detector 102 and the absorption detector 128 . label - free ) absorption and fluorescence in response to elec - 30 The memory 147 may include software and / or firmware tromagnetic radiation . With respect to the amount of amino containing instructions executed by processor 146 for con acid found in protein : Phe > Tyr > Trp . As for the extinction trolling the radiation emitting module 22 , the fluorescence coefficient for the amino acids : Trp > Tyr > Phe . The biosensor detector 102 , the absorption detector 128 , the displays 150 10 detects the amino acids specifically and defines as a and 152 , and / or other components of the biosensor 10 . The universal sensor for all biomolecules and biological cells . 35 processor 146 may convert the fluorescence signal from the

When configured as an illustrative embodiment universal fluorescence detector 102 into a fluorescence reading on the biosensor 10 for detecting both Trp and Tyr , the laser diode fluorescence display 150 . Similarly , the processor 146 may 36 illustratively emits an electromagnetic radiation or light convert the absorption signal from the absorption detector having a wavelength of between about 265 nm and about 128 into an absorbance reading on the absorption display 285 nm for the absorption measurement by the absorption 40 152 . The memory 147 may include a random - access detector 128 . In one such illustrative embodiment , the laser memory configured to store information such as the date , diode 36 emits light having a wavelength of about 275 nm . locations , and the number of positive hits for particular An illustrative laser diode has an 8 to 10 mW output power , biomolecules based upon the electromagnetic radiation emitting a wavelength of 270 to 280 nm , such as model wavelengths sensed by the detectors 102 and 128 . An UVCLEAN275SMD available from Photonics of Ann 45 electrical coupler , for example a communication port or Arbor , Mich . transmitter ( not shown ) , may be operably coupled to the

The fluorescence measurement may illustratively be set controller 145 for providing electrical communication with between 295 nm and 420 nm , primarily for Trp . In one such the processor 146 to supply data to external devices , for illustrative embodiment , this is set at 345 nm through the example . fluorescence band - pass filter 106 . 50 The display module 28 is illustratively positioned distal of

In the case for detecting Phe , the laser diode 36 illustra - the absorption detector module 26 and includes a display tively emits electromagnetic radiation of about 255 nm for module housing 148 . Fluorescence display 150 is supported the absorption measurement and excitation , while about 280 within the display module housing 148 and is configured to nm is used for the fluorescence measurement . provide an indication of the fluorescence spectral data from

As further detailed herein , the absorption detector 128 and 55 the processor 146 as received from the fluorescence detector the fluorescence detector 102 are illustratively deep ultra - 102 . Absorption display 152 is supported within the display violet ( DUV ) detectors . In alternative illustrative embodi module housing 148 and is configured to provide an indi ments , the absorption detector 128 and the fluorescence cation of the absorption spectral data from the processor 146 detector 102 may each be a charge - coupled device ( CCD ) as as received from the absorption detector 128 . The fluores used in camera and imaging devices . 60 cence display 150 illustratively comprises a digital readout When the illustrative biosensor 10 is configured as a ( e . g . , a liquid crystal display ( LCD ) ) configured to display

selective sensor for only particular biomolecules , the laser fluorescence spectral data from the processor 146 as diode 36 is selected with a suitable emission wavelength for received from the fluorescence detector 102 . Similarly , the detecting that biomolecule of interest . More particularly , in absorption display 152 illustratively comprises a digital such a configuration , the biosensor 10 demonstrates 65 readout ( e . g . , a liquid crystal display ( LCD ) ) configured to increased selectivity and sensitivity for detection . As further display absorption data from the processor 146 as received detailed herein , the biomolecule of interest in the sample from the absorption detector 128 .

US 10 , 036 , 703 B1

Different colors displayed by the display screens 150 and With reference to FIGS . 6A - 6C , an illustrative method of 152 ( e . g . , backlighting ) may indicate the detection result operating the biosensor 10 of the present disclosure is from the target sample solution in the sample cell chamber disclosed . Initially , the biosensor 10 is oriented in an upright 40 . For example , green may represent no detectable signal position with the longitudinal axis 14 extending in a vertical ( i . e . , safe ) , yellow may represent a low detection signal ( i . e . , 5 direction with the proximal end 16 positioned below the warning ) , and red may represent a significant detection distal end 18 . A waste adapter including tubing 80 and waste signal ( i . e . , alarming ) . Illustratively , the intensity level of the collection chamber or vial 82 is fluidly coupled to the sample display screens 150 and 152 are digital scales from 0 to 100 , cell chamber 40 . More particularly , an end of the tubing 80 with threshold readings representing safe , warning , and is fluidly coupled to the waste outlet 62 . alarming detection from the target sample . 10 Next , a cleansing solution ( e . g . , an alcohol - water mixture )

Illustratively , the display module housing 148 includes a of volume excess to ( i . e . , greater than ) the volume of the cylindrical side wall 154 extending between a proximal end sample cell chamber 40 is received within the barrel 76 of 156 and a distal end 158 . Illustratively , an end cap 160 is the syringe 72 . The tip 70 of the syringe 72 is received threadably coupled to the proximal end 156 of the side wall within the septum 66 of the sample inlet 58 and the cleansing 154 . Openings 162 , 164 are formed within the side wall 154 15 solution is injected into the sample cell chamber 40 by and are aligned with the displays 150 , 152 . The display depressing the plunger 74 of the syringe 72 . The excess module 28 is in electrical communication with the processor cleansing solution flows out through the waste outlet 62 and 146 . The displays 150 , 152 and the processor 146 are into the collection vial 82 . illustratively coupled to a support , such as a printed circuit The method continues by inverting the biosensor 10 to an board ( pcb ) 166 . 20 inverted position such that the longitudinal axis 14 extends

The display module 28 illustratively further includes a in a vertical direction with the distal end 18 positioned below second or distal power source 168 in electrical communi - the proximal end 16 . Air is received within the barrel 76 by cation with the processor 146 , the displays 150 , 152 , the pulling the plunger 74 of the syringe 72 with the tip 70 absorption detector 128 , and the fluorescence detector 102 . engaged . A filter 178 is then attached between the syringe 72 The second power source 168 illustratively comprises a 25 and the tip 70 . The tip 70 of the syringe 72 is received within distal battery 169 releasably supported within the end cap the septum 66 of the sample inlet 58 . Next , the plunger 74 160 . The distal battery 169 is illustratively a rechargeable is depressed thereby injecting filtered air into the sample cell button cell battery . In one illustrative embodiment , a military chamber 40 . The sample cell chamber 40 is evacuated by air grade 3 . 0 Volt Saft BA5367U Lithium Sulfur Dioxide ( 1 forcing the residual cleansing solution out through the waste inchx1 inchx0 . 075 inch ) battery produced by Saft SA in 30 outlet 62 and into the waste vial 82 , thereby helping to dry France may be used . Another illustrative battery is the the inside surface of the sample cell chamber 40 . BA53670 battery together with a DC / DC High Voltage In the next step of the method , the biosensor 10 is returned DBC - Series ( 0 . 83 inchx0 . 83 inchx0 . 32 inch ) Module avail to the upright position with the longitudinal axis 14 extend able from Laser Components USA ) for raising the voltage ing in a vertical direction with the proximal end 16 posi output . 35 tioned below the distal end 18 . A blank solution ( e . g . , water

A second or distal power switch 170 is illustratively in solution ) is received within the barrel 76 of the syringe 72 . electrical communication with the distal battery 169 and is The tip 70 of the syringe 72 is received within the septum 66 supported by the end cap 160 proximate the distal end 18 of of the sample inlet 58 . Next , the plunger 74 is depressed the outer casing 12 . The distal power switch 170 may be a thereby injecting the blank into the sample cell chamber 40 . toggle push button that is configured to be depressed by a 40 The processor 146 will electronically generate zero absor user . bance at the absorption display 152 and zero signal at the

A third releasable coupling 172 is illustratively positioned fluorescence display 150 . In one illustrative method , a between the display module 28 and the absorption detector sample solution is next received within the barrel 76 of the module 26 . Illustratively , internal threads 174 on the distal syringe 72 . The tip 70 of the syringe 72 is received within end 126 of the absorption detector module 26 engage 45 the septum 66 of the sample inlet 58 . Next , the plunger 74 external threads 176 on the proximal end 156 of the display is depressed thereby injecting the sample solution into the module 28 . Other releasable couplings may be substituted sample cell chamber 40 . for cooperating threads , including , for example bayonet In another illustrative method , a filtered sample solution couplings , resilient fingers , lock washers , etc . is received within the barrel 76 by pulling the plunger 74

Wires may electrically couple the fluorescence detector , 50 with a filter 178 attached between the syringe 72 and the tip the processor 146 ( and thereby the fluorescence display ) , 70 received in the sample solution . The filter 178 is then and the distal battery . In certain illustrative embodiments , detached . The tip 70 of the syringe 72 is received within the the releasable couplings 114 , 140 , 172 may include electrical septum 66 of the sample inlet 58 . Next , the plunger 74 is couplers , such as spring contacts to provide electrical com - depressed thereby injecting the filtered sample solution into munication between the components in the radiation emit - 55 the sample cell chamber 40 . ting module 22 , the sample cell module 24 , the absorption The blank solution or the sample solution is illustratively detector module 26 , and the display module 28 . injected into the sample cell chamber 40 within the sample

The modules 22 , 24 , 26 , 28 are illustratively fitted or inlet 58 to a volume level such that an excess volume of the assembled into the functional form of the biosensor 10 . sample solution comes out of waste outlet 62 . More particularly , in the illustrative embodiment biosensor 60 The method continues by the user depressing the power 10 , the radiation emitting module housing 30 , the sample buttons 52 , 170 to " on ” positions , thereby activating the cell chamber 40 , the sample cell module housing 54 , the radiation emitting module 22 , the display module 28 , the absorption detection module housing 120 and the display fluorescence detector 102 and the absorption detector 128 . module housing 148 cooperate to define the cylindrical outer The laser diode 36 emits electromagnetic radiation , illustra casing 12 . The releasable couplings 114 , 140 , 172 allow the 65 tively light ( e . g . , laser beam 38 ) , through the entry port 96 modules 22 , 24 , 26 , 28 to be separable , thereby facilitating and the sample chamber 40 . The light passes through the the easy removal and replacement of components . sample solution in the sample cell chamber 40 , where the

US 10 , 036 , 703 B1 13 14

absorption portion of the light ( i . e . , intensity of the laser Additionally , the occurrence of false or incorrect results is beam 38 absorbed by the sample solution ) is directed to the easily identifiable . In addition to potential molecular spectral exit port 98 and to the absorption detector 128 , and the interference , the absorption measurement may receive inter fluorescence portion of the light ( i . e . , intensity of excited or ference by beam - blockage from opaque particles and the fluorescence emissions from the sample solution as a result 5 fluorescence ( emission ) measurement may receive interfer of the laser beam 38 ) is reflected by the reflective surface ence by beam - scattering from reflective particles . Receiving 108 at a right angle to the electromagnetic radiation laser an absorption reading but not a fluorescence reading , or vice beam 38 ( i . e . , the longitudinal axis 14 ) to the fluorescence versa , may be an indication of a possible false / incorrect detector 102 result . Therefore , having both an absorption reading and a Absorption values from the absorption detector are trans - 10 fluorescence reading works as a double - assurance , thereby mitted to the processor 146 and then displayed on the increasing the confidence on each result of the biosensor . absorption display 152 . Fluorescence values from the fluo Native absorption and fluorescence provides a major rescence detector are transmitted to the processor 146 and then displayed on the fluorescence display 150 . A baseline advantage for biomolecule detection because all biomol value can be set with the blank solution , where the display 15 ecules contain an 15 ecules contain amino acids ( e . g . , phenylalanine , tryptophan , readings can be adjusted to zero values for background and tyrosine ) and , therefore , by laser or appropriate light subtraction via the processor 146 . Positive reading valves exposure directly on the sample solution followed by simul above zero indicate presence of biomolecules . The process taneous positive absorption detection and positive fluores may then continue by returning to the step of injecting the cence detection indicate the presence of biomolecules ( e . g . , cleansing solution via the syringe 72 . 20 phenylalanine , tryptophan , and tyrosine ) . That is , tryptophan As detailed above the outer dimensions of the biosensor and tyrosine characteristically absorb the laser or light

10 are relatively small . More particularly , the biosensor 10 wavelength of 275 nm and , when excited , emit their fluo is of a small size and is light weight , similar to those of a rescence radiation at 345 nm . pen / laser pointer . The biosensor 10 also includes the capa Additionally , operation of the biosensor 10 of the present bility to provide both absorption and fluorescence measure - 25 disclosure provides for improved assay time . Simply acti ments simultaneously in of a field acquired sample solution . vate the power buttons and read the displays which provide The biosensor 10 may find utility in a variety of applications , instance results in the field . The components of the biosensor including in the military , food and health industries , and are relatively inexpensive , thereby providing for a cost low with first responders . enough for wide distribution .

As further detailed herein , the biosensor 10 of the present 30 The modular design of the biosensor 10 provides com disclosure facilitates portability by having a small size , ponent choices . Assembling particular modules making the being light weight , being durable , having low power require biosensor 10 functions as a universal biosensor for all ments and being pocket wearable . The biosensor 10 includes biomolecules ( based on native molecular absorption spec modular components thereby allowing choices of integra trophotometry and native fluorescence spectrophotometry ) tion into a variety of different purpose specific biosensors . 35 or as a selective biosensor ( based on specific - tagging of

The biosensor 10 of the present disclosure provides for disclosure provides for selective biomolecules followed by characteristic absorption improved selectivity . More particularly , the biosensor 10 and fluorescence from the tagged biomolecules ) . provides for selective detection based on combined molecu As detailed herein , theoretically , all biomolecules are lar absorption spectrophotometry ( MAS ) and molecular composed an amount of amino acids tryptophan and tyro fluorescence spectrophotometry ( MFS ) . A higher signal 40 sine . Spectrally , tryptophan and tyrosine uniquely absorb reading is obtainable , due in part to the long path of laser light energy of approximately 275 nm wavelength , get beam - sample interactive , either from absorption detection or excited and subsequently emit their characteristic fluores fluorescence detection , compared to “ solo ” operation ( i . e . , cence of approximately 345 nm wavelength . Operatively as absorption detection or fluorescence detection alone ) . Some a biosensor , the laser diode 36 emits an electromagnetic molecules favor the absorption process and other molecules 45 radiation ( e . g . , laser beam 38 ) of approximately 275 nm favor the fluorescence process , due to either spectral or wavelength that is directed to pass through a sample in the fundamental interactions , generating a higher combined sample cell chamber 40 . The absorption detector 128 senses signal . the intensity of the laser beam 38 ( 275 nm wavelength )

The biosensor 10 of the present disclosure also provides absorbed , and the fluorescence detector 102 detects the for improved sensitivity . More particularly , the biosensor 10 50 collected light intensity , specifically of approximately 345 provides for high sensitivity from the sample cell including nm wavelength , emitted from the sample solution . A positive a long absorption path length and a long emission ( fluores detection ( signal above background from a blank solution ) cence ) region . Additionally , the biosensor 10 operates with from the detectors 102 and 128 indicates the sample solution a laser beam , sample cell , and detectors enclosed in a light contains biomolecules . sealed assembly to prevent the detectors receiving interfer - 55 Although the invention has been described in detail with ence from external light sources . Further , the sample cell of reference to certain preferred embodiments , variations and the biosensor may be recalibrated ( i . e . , zeroed ) by using a modifications exist within the spirit and scope of the inven blank solution or empty sample cell chamber , resulting in a tion as described and defined in the following claims . higher signal - to - noise ratio , thereby increasing the sensitiv ity of the biosensor 10 . 60 The invention claimed is :

Further , the biosensor 10 is easy to operate with minimal 1 . A hand - held biosensor comprising : or no sample preparation required . Simply inject an excess a radiation emitting module including a radiation emitting volume of a sample , and activate the power buttons to module housing , a light source received within the operate the biosensor 10 . radiation emitting module housing and configured to

The biosensor 10 demonstrates reliability by providing 65 generate electromagnetic radiation ; simple to read displays , providing repeatable performance a sample cell module including a sample cell module with no operator interpretation required in reading results . housing defining a chamber configured to receive a

US 10 , 036 , 703 B1 15 16

solution , a sample inlet in fluid communication with the releasable coupling includes a threaded connection between chamber , and a waste outlet in fluid communication the absorption detection module housing and the sample cell with the chamber ; module housing .

a first releasable coupling between the radiation emitting 10 . The hand - held biosensor of claim 1 , wherein the module and the sample cell module ; 5 sample cell module includes a light entry port , a light exit

an absorption detector configured to detect emission port axially spaced by the chamber from the light entry port , intensity produced by the electromagnetic radiation a longitudinal axis defined by the light entry port and the passing through the solution in the sample cell chamber light exit port , and a reflective surface configured to reflect and produce absorption spectral data in response light transverse to the longitudinal axis . thereto , the absorption detector including an absorption 10 11 . The hand - held biosensor of claim 10 , wherein the detector spectral filter ; sample cell module includes an axial length between the

a fluorescence detector configured to detect molecular light entry port and the light exit port , the axial length being emissions produced by the electromagnetic radiation at least 5 centimeters . passing through the solution in the sample cell chamber 12 . The hand - held biosensor of claim 10 , wherein the and produce fluorescence spectral data in response 15 sample cell is a quartz sample cell . thereto , the fluorescence detector including a fluores - 13 . The hand - held biosensor of claim 1 , wherein the cence detector spectral filter ; sample inlet comprises a resilient septum configured to

a processor in electrical communication with the absorp receive a sample syringe . tion detector and the fluorescence detector , the proces 14 . A hand - held biosensor comprising : sor configured to receive absorption spectral data from 20 an outer casing extending between a proximal end and a the absorption detector and fluorescence spectral data distal end ; from the fluorescence detector ; a radiation emitting module supported proximate the

a display module in electrical communication with the proximal end of the outer casing ; processor , the display module including a display mod a display module supported proximate the distal end of ule housing , an absorption display supported within the 25 the outer casing ; display module housing and configured to provide an a sample cell module supported intermediate the radiation indication of the absorption spectral data from the emitting module and the display module ; absorption detector , and a fluorescence display sup the sample cell module including a sample cell module ported within the display module housing and config housing defining a chamber and including a light entry ured to provide an indication of the fluorescence spec - 30 port , a light exit port axially spaced by the chamber tral data from the fluorescence detector ; from the light entry port , a longitudinal axis defined by

a power source in electrical communication with the the light entry port and the light exit port , and a processor ; and reflective surface configured to reflect light transverse

a second releasable coupling between the display module to the longitudinal axis ; and the sample cell module . 35 an absorption detector ;

2 . The hand - held biosensor of claim 1 , wherein the light a fluorescence detector ; source comprises a laser diode . a processor in electrical communication with the absorp

3 . The hand - held biosensor of claim 1 , further comprising tion detector and the fluorescence detector , the proces an absorption detection module including an absorption sor configured to receive absorption spectral data from detection module housing receiving the absorption detector , 40 the absorption detector and fluorescence spectral data and a third releasable coupling between the absorption from the fluorescence detector ; and detection module and the sample cell module , wherein the wherein the sample cell module includes an axial length second releasable coupling is between the display module between the light entry port and the light exit port , the and the absorption detection module . axial length being at least 5 centimeters .

4 . The hand - held biosensor of claim 3 , wherein the 45 15 . The hand - held biosensor of claim 14 , wherein : radiation emitting module housing , the display module the radiation emitting module includes a radiation emit housing , and the absorption detection module housing coop ting module housing , a light source received within the erate to define a cylindrical outer casing . radiation emitting module housing and configured to

5 . The hand - held biosensor of claim 4 , wherein the generate an excitation energy ; and cylindrical outer casing extends between a proximal end and 50 the display module includes a display module housing , an a distal end , the outer casing having an axial length of about absorption display supported by the display module 20 centimeters and a diameter of about 4 centimeters . housing and configured to provide an indication of the

6 . The hand - held biosensor of claim 5 , further comprising absorption spectral data , and a fluorescence display an external clip coupled proximate the distal end of the outer supported by the display module housing and config casing . 55 ured to provide an indication of the fluorescence spec

7 . The hand - held biosensor of claim 5 , wherein the power tral data . source includes a proximal battery and a power switch 16 . The hand - held biosensor of claim 15 , further com positioned proximate the proximal end of the outer casing . prising :

8 . The hand - held biosensor of claim 7 , wherein the power a first releasable coupling between the radiation emitting source includes a distal battery and a power switch posi - 60 module and the sample cell module ; and tioned proximate the distal end of the outer casing . a second releasable coupling between the display module

9 . The hand - held biosensor of claim 3 , wherein the first and the sample cell module . releasable coupling includes a threaded connection between 17 . The hand - held biosensor of claim 16 , further com the radiation emitting module housing and the sample cell prising an absorption detection module including an absorp module housing , the second releasable coupling includes a 65 tion detection module housing receiving the absorption threaded connection between the display module housing detector , and a third releasable coupling between the absorp and the absorption detection module housing , and the third tion detection module and the sample cell module , wherein

US 10 , 036 , 703 B1 18

the second releasable coupling is between the display mod and a fluorescence display supported within the display ule and the absorption detection module . module housing and configured to provide an indica

18 . The hand - held biosensor of claim 17 , wherein the first tion of the fluorescence spectral data ; releasable coupling includes a threaded connection between releasably coupling the distal end of the sample cell the radiation emitting module housing and the sample cell 5 module housing to the proximal end of the display module housing , the second releasable coupling includes a module housing ; and threaded connection between the display module housing providing a power source in electrical communication and the absorption detection module housing , and the third with the processor . releasable coupling includes a threaded connection between 24 . The method of claim 23 , wherein the light source the absorption detection module housing and the sample cell 10 comprises a laser diode and a laser diode driver operably module housing .

19 . The hand - held biosensor of claim 18 , wherein the coupled to the laser diode , the laser diode configured to radiation emitting module housing , the sample cell module project light to a lens . housing , the display module housing , and the absorption 25 . The method of claim 23 , wherein the steps of : detection module housing cooperate to define the outer 15 5 releasably coupling the distal end of the radiation emitting casing . module housing to the proximal end of the sample cell

20 . The hand - held biosensor of claim 19 , wherein the module housing includes threadably coupling the radia outer casing is cylindrical and extends between a proximal tion emitting module housing with the sample cell end and a distal end , the outer casing having an axial length module housing ; and of about 20 centimeters and a diameter of about 4 centime - 20 releasably coupling the distal end of the sample cell ters . module housing to the proximal end of the display

21 . The hand - held biosensor of claim 14 , wherein the module housing includes threadably coupling the sample cell module includes an axial length between the sample cell module housing with the display module light entry port and the light exit port , the axial length being housing . at least 5 centimeters . 25 26 . The method of claim 23 , further comprising the steps

22 . The hand - held biosensor of claim 21 , wherein the of : sample cell is a quartz sample cell . providing an absorption detection module including an

23 . A method of constructing a hand - held biosensor , the absorption detection module housing extending longi method comprising the steps of : tudinally between a proximal end and a distal end and

providing a sample cell module including a sample cell 30 receiving the absorption detector ; module housing having a side wall defining a chamber releasably coupling the distal end of the sample cell and extending longitudinally between a proximal end module housing with the proximal end of the absorp and a distal end , a sample inlet in fluid communication tion detection module housing ; and with the chamber , and a waste outlet in fluid commu releasably coupling the distal end of the absorption detec nication with the chamber ; 35 tion module housing module and the proximal end of

providing a solution within the chamber of the sample cell the display module housing . module ; 27 . The method of claim 26 , wherein :

providing a radiation emitting module including a radia the radiation emitting module housing , the sample cell tion emitting module housing extending longitudinally module housing , the display module housing , and the between a proximal end and a distal end , a light source 40 absorption detection module housing cooperate to received within the radiation emitting module housing define a cylindrical outer casing ; and the outer casing and configured to generate electromagnetic radiation ; extends between a proximal end and a distal end , the

releasably coupling the distal end of the radiation emitting outer casing having an axial length of about 20 centi module housing to the proximal end of the sample cell meters and a diameter of about 4 centimeters . module housing ; 45 28 . The method of claim 23 , wherein the sample cell

providing an absorption detector at the distal end of the module includes a light entry port , a light exit port axially sample cell , the absorption detector configured to spaced by the chamber from the light entry port , a longitu detect emission intensity produced by the electromag - dinal axis defined by the light entry port and the light exit netic radiation passing through the solution in the port , and a reflective surface configured to reflect light sample cell chamber , the absorption detector including 50 transverse to the longitudinal axis . an absorption detector spectral filter ; 29 . The method of claim 28 , wherein the sample cell

providing a fluorescence detector configured to detect module includes an axial length between the light entry port molecular emissions produced by the electromagnetic and the light exit port , the axial length being at least 5 radiation passing through the solution in the sample cell centimeters . chamber , the fluorescence detector including a fluores - 55 30 . A method of detecting a substance within a fluid cence detector spectral filter ; sample using a hand - held biosensor , the method comprising

providing a processor in electrical communication with the steps of : the absorption detector and the fluorescence detector , providing a sample cell module including a sample cell the processor configured to receive absorption spectral module housing having a side wall defining a chamber data from the absorption detector and fluorescence 60 and extending a longitudinal axis between a proximal spectral data from the fluorescence detector ; end and a distal end , a sample inlet in fluid communi

providing a display module in electrical communication cation with the chamber , and a waste outlet in fluid with the processor , the display module including a communication with the chamber ; display module housing extending between a proximal fluidly coupling a waste collection vial to the waste outlet ; end and a distal end , an absorption display supported 65 injecting a sample solution through the sample inlet into within the display module housing and configured to the chamber of the sample cell module ; provide an indication of the absorption spectral data , generating electromagnetic radiation with a light source ;

1ce11

US 10 , 036 , 703 B1 19 20

directing the electromagnetic radiation through the 37 . The method of claim 31 , wherein the light source is sample solution in the chamber of the sample cell ; received within a radiation emitting module housing of a

absorbing the electromagnetic radiation by the sample radiation emitting module , and further comprising the step solution ; of releasably coupling the radiation emitting module hous sensing the intensity of the electromagnetic radiation 5 5 ing to the sample cell module housing . absorbed by the sample solution by an absorption 38 . The method of claim 37 , wherein the absorption detector ;

generating an absorption signal of interest from the display and the fluorescence display are received within a absorption detector ; display module housing of a display module , and further

producing fluorescence emissions from the sample solu - 10 comprising the step of releasably coupling the display tion ; module housing to the sample cell module housing .

sensing the fluorescence emissions by a fluorescence 39 . The method of claim 31 , wherein the sample cell detector ; module includes a light entry port , a light exit port axially generating a fluorescence signal of interest from the spaced by the chamber from the light entry port , a longitu fluorescence detector ; dinal axis defined by the light entry port and the light exit displaying an indication of the absorption signal on an 13 port , and a reflective surface configured to reflect light absorption display ; and

displaying an indication of the fluorescence signal on a transverse to the longitudinal axis . tran fluorescence display . 40 . The method of claim 39 , wherein the sample cell

31 . The method of claim 30 , further comprising the step module includes an axial length between the light entry port of injecting a cleansing solution through the sample inlet 20 and the light exit port , the axial length being at least 5 prior to the step of injecting the sample solution . centimeters .

32 . The method of claim 31 , further comprising the steps 41 . The method of claim 30 , wherein the light source of inverting the sample cell , and injecting filtered air through generates electromagnetic radiation for the sample solution the sample inlet port , and forcing the cleansing solution out having a wavelength of approximately 275 nm , and fluo through the waste outlet . 25 rescence emissions from the sample solution having a wave 33 . The method of claim 30 , further comprising the step length of approximately 345 nm . of contacting a first power user input to activate the light 42 . The method of claim 41 , wherein the light source source .

34 . The method of claim 33 , further comprising the step comprises a laser diode and a laser diode driver operably of contacting a second power user input to activate the 30 coupled to the laser diode , the laser diode configured to absorption detector , the fluorescence detector , the absorption O project light to a lens .

43 . The method of claim 41 , further comprising a band display and the fluorescence display . 35 . The method of claim 31 , wherein the step of directing pass spectral filter to generate the selected fluorescence |

the excitation energy includes : emissions having a wavelength of approximately 345 nm . directing light parallel to a longitudinal axis through the 35 44 . The method of claim 41 , wherein the electromagnetic

chamber of the sample cell to the absorption detector ; 35 radiation having a wavelength of approximately 275 nm is and absorbed by amino acids including tryptophan and tyrosine .

45 . The method of claim 44 , wherein tryptophan and reflecting light transverse to the longitudinal axis through the chamber of the sample cell to the fluorescence tyrosine are excited on absorption of radiation having a

wavelength of 275 nm . detector . 36 . The method of claim 31 , wherein the sample inlet + 40 46 . The method of claim 45 , wherein tryptophan and

includes a septum , and the step of injecting a sample tyrosine emit fluorescence emissions having a wavelength of 345 nm . solution through the sample inlet includes inserting a syringe

into the septum . * * * * *