IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
`US008933395B2
`
`(12) United States Patent
`Mueth et al.
`
`(10) Patent No. :
`(45) Date of Patent:
`
`US 8,933,395 B2
`Jan. 13, 2015
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`3,390,449 A
`3,649,829 A
`
`7/1968 Fox
`3/1972 Randolph
`
`(Continued)
`
`FOREIGN PATENT DOCUMENTS
`
`DE
`EP
`
`19952322 Al
`0057907 Al
`(Continued)
`
`5/2001
`8/1982
`
`OTHER PUBLICATIONS
`
`Hori M, et al. , "Cell fusion by optical
`trapping with laser
`involves
`. ", WPI/Thomson, Dec. 27, 1991,
`different cells.
`contacting
`.
`Abstract.
`
`(Continued)
`
`Kiet T Nguyen
`Primary Examiner
`Jean C. Edwards, Esq. ;
`(74) Attorney, Agent, or Firm
`Edwards Neils PLLC
`
`ABSTRACT
`(57)
`The present
`to
`a method and apparatus
`invention provides
`from a plurality of compo-
`identify at least one component
`the apparatus
`nents in a fluid mixture,
`including a first input
`into which a first flow containing a plurality of com-
`channel
`a plurality of buffer
`is introduced;
`input channels,
`ponents
`flows of buffer solution are introduced,
`into which additional
`disposed on either side of the first input channel; wherein the
`flows have a flow direction along
`first flow and the additional
`a length of the apparatus;
`a detector apparatus which detects
`selected components of the plurality of com-
`and identifies
`ponents; a laser which emits a laser beam which damages or
`kills selected components ofthe plurality of components;
`and
`at least one channel disposed at the another end of the appa-
`ratus which is adapted to receive the first flow and the addi-
`flows after operation of the laser on the selected com-
`tional
`ponents.
`
`14 Claims, 22 Drawing Sheets
`
`(72)
`
`Inventors:
`
`(73) Assignee:
`
`( * ) Notice:
`
`(54) MULTIPLE LAMINAR FLOW-BASED
`PARTICLE AND CELLULAR
`IDENTIFICATION
`Premium Genetics (UK) Ltd. ,
`(71) Applicant:
`Nantwich (GB)
`Daniel Mueth, Chicago, IL (US);
`Joseph Plewa, Park Ridge, IL (US);
`Jessica Shireman, Kansas City, MO
`IL (US);
`(US); Amy Anderson, Palatine,
`Lewis Gruber, Chicago, IL (US); Neil
`Rosenbaum, Chicago, IL (US)
`Premium Genetics (UK) Ltd. , Cheshire
`(GB)
`the term of this
`Subject to any disclaimer,
`is extended or adjusted under 35
`patent
`U.S.C. 154(b) by 0 days.
`14/169,927
`Jan. 31, 2014
`Prior Publication Data
`May 29, 2014
`
`(21) Appl. No. :
`Filed:
`(22)
`(65)
`
`US 2014/0147881 Al
`
`Related U.S.Application Data
`(60) Continuation of application No. 13/412, 969, filed on
`Mar. 6, 2012, now Pat. No. 8,653,442, which is a
`(Continued)
`
`(51)
`
`Int. Cl.
`B01D 21/26
`G02B 21/32
`
`(2006.01)
`(2006.01)
`(Continued)
`
`(52) U.S. Cl.
`CPC ............ G01N 33/48 7 (2013.01); G03H 1/2294
`(2013.01); G03H 2001/0077 (2013.01);
`(Continued)
`(58) Field of ClassiTication Search
`USPC ............... 250/251; 356/244, 246; 494/36, 45;
`435/173. 1; 210/732, 800, 802
`See application file for complete search history.
`
`HN. MIl/@at
`OPIIt/L IRIIPPING
`IIII
`
`

`

`US 8,933,395 B2
`Page 2
`
`Related U.S.Application Data
`continuation of application No. 12/659, 277, filed on
`Mar. 2, 2010, now Pat. No. 8, 158,927, which is a
`division of application No. 12/213, 109, filed on Jun.
`13, 2008, now Pat. No. 7,699,767, which is a division
`of application No. 11/543,773, filed on Oct. 6, 2006,
`now Pat. No. 7,402, 131, which is a division of appli-
`cation No. 10/934, 597, filed on Sep. 3, 2004, now Pat.
`of
`No. 7,118,676, which
`is a continuation-in-part
`application No. 10/867, 328, filed on Jun. 13, 2004,
`now Pat. No. 7, 150,834, which is a continuation-in-
`part of application No. 10/630, 904, filed on Jul. 31,
`2003, now Pat. No. 7,241,988.
`application No. 60/399, 386, filed on Jul.
`Provisional
`31, 2002, provisional
`application No. 60/435, 541,
`filed on Dec. 20, 2002, provisional
`application No.
`60/571, 141, filed on May 14, 2004, provisional
`appli-
`cation No. 60/499, 957, filed on Sep. 4, 2003, provi-
`sional application No. 60/511,458, filed on Oct. 15,
`2003.
`
`(60)
`
`(51)
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`Int. Cl.
`G01N 33/48 7
`B01D 21/01
`H05H 3/04
`G03H 1/22
`G03H 1/00
`G03H 1/08
`(52) U.S. Cl.
`CPC ................ G02B21/32 (2013.01);B01D 21/01
`(2013.01); G03H 2225/32 (2013.01); G03H
`1/08 (2013.01); H05H 3/04 (2013.01); G03H
`2001/085 (2013.01)
`USPC ............. 250/251; 356/244; 356/246; 494/36;
`494/45; 435/173. 1; 210/732; 210/800; 210/802
`
`... 356/244
`
`6,416,190 Bl
`6,432,630 Bl
`6,451,264 Bl
`6,506,609 Bl
`6,524, 860 Bl
`6,637,463 Bl
`6,727,451 Bl
`6,833,542 B2
`6,838,056 B2
`6,944,324 B2
`7,029,430 B2
`7,241,9gg B2
`7,355,696 B2 *
`7,472,794 B2
`7,482, 577 B2
`8, 158,122 B2
`2002/0058332 Al
`2002/0176069 Al
`2003/0032204 Al
`2003/0047676 Al
`2003/0186426 Al
`2005/0061962 Al
`2005/0121604 Al
`2006/0058167 Al
`2006/0152707 Al
`
`7/2002
`8/2002
`9/2002
`I/2003
`2/2003
`10/2003
`4/2004
`12/2004
`I/2005
`9/2005
`4/2006
`7/2007
`4/2008
`I/2009
`I/2009
`4/2012
`5/2002
`11/2002
`2/2003
`3/2003
`10/2003
`3/2005
`6/2005
`3/2006
`7/2006
`
`Grier et al.
`Blankenstein
`Bhullar et al.
`Wada et al.
`Seidel et al.
`Lei et al.
`Fuhr et al.
`Wang et al.
`Foster
`Tran et al.
`et al.
`Hlavinka
`Gruber et al.
`Mueth et al.
`Oakey et al.
`Gruber et al.
`Hampson et al.
`Quake et al.
`Hansen et al.
`Walt et al.
`Grier et al.
`Brewer et al.
`Mueth et al.
`Mueth et al.
`Ragusa et al.
`Kanda
`
`.......
`
`FOREIGN PATENT DOCUMENTS
`
`EP
`FR
`JP
`JP
`JP
`JP
`JP
`JP
`JP
`JP
`JP
`JP
`JP
`JP
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`
`0679325 Al
`2798557 Al
`57-131451 A
`58-090513 A
`H05-26799 A
`H-06-265452 A
`06-327494
`07-024309
`Hll-508182 A
`2000-512541 A
`2001-504936 A
`2002-503334 A
`2002-153260
`2005-502482 A
`97/00442 Al
`97/39338 Al
`97/47390 Al
`98/10267 Al
`99/39223 Al
`01/18400 Al
`02/087792 Al
`03/062867 Al
`2004/012133 A2
`
`11/1995
`3/2001
`8/1982
`5/1983
`2/1993
`9/1994
`11/1994
`I/1995
`7/1999
`9/2000
`4/2001
`I/2002
`5/2002
`I/2005
`I/1997
`10/1997
`12/1997
`3/1998
`8/1999
`3/2001
`11/2002
`7/2003
`2/2004
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`4,325,706 A
`4,409, 106 A
`4,424, 132 A
`4,660,971 A
`4,667,830 A
`4,919,817 A
`5,007,732 A
`5,030,002 A
`5, 100,627 A
`5, 180,065 A
`5, 194,909 A
`5,229,297 A
`5,483,469 A
`5,620,857 A
`5,674,743 A
`5,800,690 A
`5,837, 115 A
`5,849, 178 A
`5,879,625 A
`5,966,457 A
`6,053,856 A
`6,071,442 A
`6, 185,664 Bl
`H1960 H
`6,368,871 Bl
`
`4/1982
`10/1983
`I/1984
`4/1987
`5/1987
`4/1990
`4/1991
`7/1991
`3/1992
`I/1993
`3/1993
`7/1993
`I/1996
`4/1997
`10/1997
`9/1998
`11/1998
`12/1998
`3/1999
`10/1999
`4/2000
`6/2000
`2/2001
`6/2001
`4/2002
`
`Gershman et al.
`Furuta et al.
`Iriguchi
`Sage et al.
`Jr, et al.
`Nozaki,
`et al.
`Schoendorfer
`Ohki et al.
`North, Jr.
`..............
`Buican et al.
`Touge et al.
`Tycko
`Schnipelsky et al.
`Van den Engh et al.
`Weetal1 et al.
`Ulmer
`Chow et al.
`Austin et al.
`Holm et al.
`Roslaniec et al.
`Lemelson
`Hlavinka
`Dean et al.
`Jeddeloh
`Conrad et al.
`Christel et al.
`
`356/73
`
`OTHER PUBLICATIONS
`
`S. Takayama
`et al. , "Patterning cells and their environments
`using
`. . ", Proc. Natl. Acad. Sci. USA, May
`fluid flows.
`laminar
`multiple
`1999, pp. 5545-5548, vol. 96.
`Paul O.P. Ts'o, "Basic Principles
`in Nucleic Acid Chemistry",
`National Library of Medicine, 1974, pp. 311-387, Academic Press
`Inc. , New York, NY.
`Stephen P. Smith et al. , Inexpensive Optical Tweezers
`graduate Laboratories, Am. J. Phys. , Jan. 1999, vol. 67.
`Otfrce Action issued by USPTO on Jul. 17, 2014 in connection with
`related U.S. Appl. No. 14/317, 738.
`et al. , "Patterning Cells and Their Environments Using
`Takayama
`Multiple Laminar Fluid Flows in Capillary Networks. "Proceedings
`of National Academy of Sciences, USA 96 i 19991.
`Final Notice of Reasons
`for Rejection,
`issued by Japanese Patent
`Otfrce on Oct. 28, 2014, in related Japanese Patent Application No.
`2011-256171.
`* cited by examiner
`
`for Under-
`
`

`

`U.S. Patent
`
`U.S. Patent
`
`Jan. 13, 2015
`Jan.13,2015
`
`Sheet 1 of 22
`Sheet],of22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`
`
`

`

`U.S. Patent
`
`U.S. Patent
`
`Jan. 13, 2015
`Jan.13,2015
`
`Sheet 2 of 22
`Sheet20f22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`=e N6H1
`
`==nn-n---
`
`
`
`madmb4<uthw~uHmm«mm04om
`
`

`

`U.S. Patent
`
`U.S. Patent
`
`Jan. 13, 2015
`510
`
`nf03mhS
`Sheet 3 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
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`n...\..
`m.mHm
`
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`amogm
`
`mwzmzcmzou
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`

`

`U.S. Patent
`
`U.S. Patent
`
`Jan. 13, 2015
`Jan.13,2015
`
`Sheet 4 of 22
`Sheet4of22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`
`
`FIG. 4
`
`OOGODOOOOOO
`
`.OOOOOOOOOD.
`
`OOQOOOcccOfi
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`

`

`U.S. Patent
`
`U.S. Patent
`
`m.mHm
`
`Jan. 13, 2015
`
`Sheet 5 of 22
`nM5amS
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`mumemmmz
`
`mzbfiamHmomwIIEEEBAll-.1All-ix
`meflh.‘3m
`noilzllwgmfim
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`5isman.
`
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`
`538gm
`
`

`

`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 6 of 22
`Sheet 6 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`800
`
`FIG. 8A
`
`PTTOTIOI A FIRST FLGTT' HATTTTO A PLUHALITY
`(IF TOTEOTTETTTS
`
`PROVIDE A SEcOTTO FLOW
`
`EONTACT THE FIRST FLOW WITHTHE SECOND FLOW
`TO PROVIDE A FIRST SEPARATION REGION
`
`DIFFERBTTIALLY SEDIHENT A FIRST COMPQNENT
`OF A PLURALITY 0F COMPONENTS FROM THE FIRST
`FLOW INTO THE SECGNU FLOW
`
`OOTOUOOETTTIT MAINTAIN A SECGNT) TOTTTOTTITTT OF THE
`PLOTIIITT OE COKPONENTS IN THE FIRST FLUTT
`
`OTFFETTETTTIALLT BETTOTI THE SECOND ELOTT HITTING
`THE FIRST COMPONENT FOOT THE FIRST FIOTT HAVING
`THE SECOND COTTPOTTETTT
`
`505
`
`51“
`
`515
`
`539
`
`825
`
`830
`
`335
`
`.
`
`fig§3§
`
`YES
`
`“0
`0
`
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`
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`TO PROVIDE A SECOND SEPARATION REGION
`
`
`HDLOSRAFBIC
`
`HANIPULATION OFT
`SEPARATION?
`
`
`PHDVIDE A TTTTOO ELOTT
`
`5““
`
`5‘45
`
`650
`
`
`
`
`TESo
`
`

`

`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 7 of 22
`Sheet 7 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`FIG. 68
`
`emu: A PLlfiTALITY
`or momma mAPs
`
`usms m1: HOLOBHAPHIC
`TRAPS. DTFFERET‘TTIALLY MOVING
`THE SECOND COMMENT FROM
`THE FIRST FLOW INTO THE
`
`THIRO FLOW
`
`3'55
`
`880
`
`ass
`
`DIFFEHEWLLT
`SETTLEMENTS THE
`SECOND COMPONENT
`FHOH THE FIRST
`FLOW INTO THE
`
`THIRD FLOW
`
`comma MAINTABTINB A mm!) comma
`0; m5 mummy 0F moms IN THE
`FIRST How
`
`670
`
`DIFFERENTIALLY HEHOTINB THE THIRD FLOW
`HAVING THE SECOND COTTPOTTENT FROM TTE
`FIRST FLOW HAVING THE THIRD COMPONENT
`
`575
`
`RETURN
`
`BBO
`
`

`

`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 8 of 22
`Sheet 8 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`FIG.
`
`7A
`
`11'
`
`03
`
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`
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`
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`
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`
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`
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`

`

`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 9 of 22
`Sheet 9 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`FIG. 8
`
`FIG. 9
`
`
`
`
`
`FIG. 10
`
`

`

`U.S. Patent
`
`U.S. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 10 of 22
`Sheet100f22
`
`US 8,933,395 B2
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`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 11 of 22
`Sheet 11 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
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`FIG. 12
`
`1283
`
`1295
`
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`
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`
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`
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`1201
`
`1204
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`FIG. 13
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`

`U.S. Patent
`
`U.S. Patent
`
`Jan. 13, 2015
`5
`
`Sheet 12 of 22
`2
`
`US 8,933,395 B2
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`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 13 of 22
`Sheet 13 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
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`
`
`INPUT
`SfiLUTIBN
`
`
`FLWAWLU em
`‘
`summon E. E SELECTED
`PERISTALTIC Pun?
`
`man
`
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`
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`
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`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 14 of 22
`Sheet 14 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`FIG. 18
`
`um
`(63.9. CGVEB GLASS!
`
`SYRINGE
`
`DEPYH
`
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`OR TUBING
`
`1600
`
`m mm
`MATERIALS
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`
` {ABOVE EACH
`
`

`

`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 15 of 22
`Sheet 15 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`FIG. 19
`
`mm
`sowmu
`
`WASTE
`Vsuwmm
`
`BUFFER
`
`SOLUTIGN
`
`£
`
`GNE 0F w
`HIG-I-ASPECT-RATIO
`
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`
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`
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`
`
`
`
`
`
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`
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`
`
`
`
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`

`U.S. Patent
`US. Patent
`
`Jan. 13, 2015
`Jan. 13, 2015
`
`Sheet 16 of 22
`Sheet 16 of 22
`
`US 8,933,395 B2
`US 8,933,395 B2
`
`REGION DEPTH
`
`WASTE FLOW
`
`
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`FIG. 24A
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`SUPPORT LAYER
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`LAYER 2
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`INPUTS
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`FIG.27
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`US 8,933,395 B2
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`1
`MULTIPLE LAMINAR FLOW-BASED
`PARTICLE AND CELLULAR
`IDENTIFICATION
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS AND PRIORITY CLAIMS
`
`application of
`The present
`is a continuation
`application
`parent U.S. patent application Ser. No. 13/412, 969 filed Mar.
`6, 2012, now U.S.Pat. No. 8,653,442, which is a continuation
`application of U.S. patent application Ser. No. 12/659, 277,
`filed Mar. 2, 2010, now U.S. Pat. No. 8, 158,927, which is a
`application of U.S. patent
`application Ser. No.
`divisional
`12/213, 109, filed Jun. 13, 2008, now U.S.Pat. No. 7,699,767,
`application of U.S. patent application
`which is a divisional
`Ser. No. 11/543,773, filed Oct. 6, 2006, now U.S. Pat. No.
`application of U.S. patent
`7,402, 131, which is a divisional
`application Ser. No. 10/934, 597, filed Sep. 3, 2004, now U.S.
`of U.S.
`Pat. No. 7,118,676, which is a continuation-in-part
`patent application Ser. No. 10/867, 328, filed Jun. 13, 2004,
`now U.S. Pat. No. 7,150,834, which is a continuation-in-part
`of U.S. patent application Ser. No. 10/630, 904, filed Jul. 31,
`2003, now U.S. Pat. No. 7,241,988, and claims priority via
`U.S. patent application Ser. No. 10/630, 904, to U.S. Provi-
`sional Patent Application No. 60/399, 386, filed Jul. 31, 2002,
`and 60/435, 541, filed Dec. 20, 2002, and claims priority via
`U.S. patent application Ser. No. 10/934, 597, to U.S. Provi-
`sional Patent Application No. 60/571, 141, filed May 14,
`2004, U.S. Provisional Patent Application No. 60/499, 957,
`filed Sep. 4, 2003, and U.S. Provisional Patent Application
`No. 60/511, 458, filed Oct. 15, 2003, commonly
`assigned
`the contents of all of which are incorporated
`herewith,
`by
`reference herein.
`invention is related to Jessica Shireman et al. ,
`The present
`U.S. Provisional
`Ser. No. 60/571, 141,
`Patent Application
`filed May 14, 2004, entitled "System and Method of Sorting
`Blood Cells Using Holographic Laser Steering", commonly
`the contents of which are incorporated by
`assigned herewith,
`reference herein, with priority claimed for all commonly dis-
`(the "second related application" ).
`closed subject matter
`invention is related to and a conversion to a full
`The present
`utility application of Daniel M. Mueth, U.S. Patent Applica-
`tion Ser. No. 60/499, 957, filed Sep. 4, 2003, entitled "Passive
`the contents of
`Fluidic Sorter", commonly assigned herewith,
`by reference herein, with priority
`which are incorporated
`claimed for all commonly disclosed subject matter (the "third
`related application" ).
`invention is related to and a conversion to a full
`The present
`utility application of Daniel M. Mueth, U.S. Patent Applica-
`tion Ser. No. 60/511, 458, filed Oct. 15, 2003, entitled "Pas-
`sive Fluidic Sorter", commonly assigned herewith,
`the con-
`tents of which are incorporated
`by reference herein, with
`priority claimed for all commonly disclosed subject matter
`(the "fourth related application" ).
`invention is related to Lewis Gruber et al. , U.S.
`The present
`patent application Ser. No. 10/630, 904, filed Jul. 31, 2003,
`entitled "System and Method of Sorting Materials Using
`Holographic Laser Steering", commonly assigned herewith,
`the contents of which are incorporated by reference herein,
`claimed for all commonly
`disclosed subject
`with priority
`(the "fifth related application" ).
`matter
`
`FIELD OF THE INVENTION
`
`The present
`invention relates generally to techniques
`and
`for separation of particulate
`or cellular materials
`systems
`such as blood, semen and other particles or cells into their
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`and fractions, using multiple
`components
`various
`laminar
`flows which further may be coupled with laser steering such
`as holographic
`tapping and manipulation.
`optical
`
`BACKGROUND OF THE INVENTION
`
`There are several categories of blood cells. Erythrocyte or
`red blood cell (RBC) counts are for women 4.8 million cells/
`pl and men 5.4 million cells/4 RBCs make up 93%ofthe solid
`in blood and about 42% of blood volume. Platelets
`element
`are 2 pm-3 pm in size. They represent 7% of the solid ele-
`in blood and about 3% of the blood volume,
`corre-
`ments
`to about 1.5 to 4x10" cells per liter. There are 5
`sponding
`types of white blood cells (WBCs) or leukocytes
`general
`accounting for about 1.5 to 4x10 cells per liter. The WBCs
`comprise: 50-70% Neutrophils
`(12-15 pm in size); 2-4%
`(12-15 pm in size); 0.5-1% Basophils (9-10 pm
`Eosinophils
`in size); 20-40% Lymphocytes
`(25% B-cells and 75% T-cells)
`(8-10 pm in size); and 3-8% Monocytes
`(16-20 pm in size).
`They comprise 0.16% of the solid elements
`in the blood, and
`0.1% of the blood volume corresponding
`to
`approximately
`around 4 to 12x10 per liter. A subject with an infection might
`have a WBC count as high as 25x10 per liter.
`Platelets are the smallest cells in the blood and are impor-
`tant for releasing proteins
`into the blood that are involved in
`that cause lower
`clotting. Patients with immune
`diseases
`(such as cancer,
`and other chemotherapy
`counts
`leukemia
`patients) sometimes need platelet
`trans fusions to prevent
`their
`counts from becoming too low. The platelet count in adults is
`normally between 140,000-440, 000 cells/pl, and this number
`should not fall below 50,000 cells/4 because platelets play an
`role in blood clotting.
`integral
`Blood separation techniques
`have traditionally
`employed
`discrete centrifugation processes. More particularly,
`a certain
`volume of blood is removed from a donor at a particular
`time.
`That volume of blood is then subjected to different
`levels of
`for
`blood fractions
`to provide corresponding
`centrifugation
`red blood cells,
`blood components
`such as plasma, platelets,
`and white blood cells. This process is discrete,
`rather
`than
`such that if more blood from the donor is to be
`continuous,
`another volume is removed from the donor, and
`processed,
`the process is repeated.
`in platelet collection are: collection of blood
`The steps
`from donor: addition of anticoagulant;
`separation via cen-
`return of red cells, leukocytes and plasma to the
`trifugation;
`contains about 200-400 ml of
`donor. A collection normally
`plasma, which is reduced to avoid incompatibility. This col-
`about 8 to 8.5x10' platelets. A
`lection normally
`contains
`10% of his/her platelets
`donor normally gives approximately
`with no loss in clotting ability, although a larger number of
`platelets could be separated from the blood. These platelets
`must be used within five days of collection.
`is a state of the art pro-
`called apheresis,
`Plateletpheresis,
`cess by which platelets are separated [Haemonetics Compo-
`nent Collection System (CCS) and Multi Component System
`Braintree, Mass. )]. This automated
`(Multi)(Haemonetics,
`machine separates platelets from blood over a period of 1.5 to
`2 hours (assuming 10% donation). This process is faster than
`and is completely automated and can
`traditional
`approaches
`be used for single or double platelet doses. Nevertheless,
`the
`to the patience of donors
`and is
`process
`is slow relative
`capable of improvement
`for the purity of the separated plate-
`let fraction.
`often taking
`are also time consuming,
`Other procedures
`several hours, particularly when unused blood fractions are to
`be returned to the donor. For example, platelet donation make
`take several hours, as whole blood is removed from the donor,
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`US 8,933,395 B2
`
`to obtain the platelets,
`fractionated
`centrifugation
`through
`are then injected back
`and the remaining blood components
`process is also compara-
`into the donor. This centrifugation
`in damage to a proportion of the
`tively harsh, also can result
`harvested cells, effectively reducing the usable yield of the
`blood fractions.
`Other types of separations
`are also either time consuming
`or cannot process large volumes of material
`in a timely fash-
`ion. For example,
`in which viable and motile
`sperm sorting,
`sperm are isolated from non-viable or non-motile
`is
`sperm,
`task, with severe volume restrictions.
`often a time-consuming
`As discussed below in greater detail
`in describing
`the
`of particles,
`such as that
`invention, manipulations
`present
`described in the second and fifth related applications, may
`also be part ofa novel separation technique. One conventional
`technique in manipulating microscopic obj ects is optical trap-
`ping. An accepted description of the effect of optical trapping
`tightly focused light, such as light focused by a high
`is that
`aperture microscope
`lens, has a steep intensity
`numerical
`forces of a beam of
`traps use the gradient
`gradient. Optical
`to trap a particle based on its dielectric constant.
`light
`its energy, a particle having a dielectric con-
`To minimize
`than the surrounding medium will move to the
`stant higher
`region of an optical trap where the electric field is the highest.
`Particles with at least a slight dielectric constant differential
`are sensitive to this gradient and are
`with their surroundings
`either attracted to or repelled from the point of highest
`light
`is, to or from the light beam's
`focal point. In
`that
`intensity,
`forces from a
`constructing
`an optical
`trap, optical gradient
`single beam of light are employed to manipulate
`the position
`of a dielectric particle immersed
`in a fluid medium with a
`than that of the particle, but reflect-
`refractive index smaller
`ing, absorbing and low dielectric constant particles may also
`be manipulated.
`force in an optical trap competes with
`The optical gradient
`radiation pressure which tends to displace the trapped particle
`along the beam axis. An optical trap may be placed anywhere
`within the focal volume of an objective lens by appropriately
`selecting the input beam's propagation direction and degree
`of collimation. A collimated beam entering the back aperture
`of an objective lens comes to a focus in the center of the lens'
`focal plane while another beam entering at an angle comes to
`a focus off-center. A slightly diverging beam focuses down-
`stream of the focal plane while a converging beam focuses
`upstream. Multiple beams entering the input pupil of the lens
`each form an optical trap in the focal volume
`simultaneously
`at a location determined by its angle of incidence. The holo-
`technique uses a phase modifying
`graphic optical
`trapping
`to impose the phase pattern for
`diffractive optical element
`multiple beams onto the wavefront of a single input beam,
`the single beam into multiple
`traps.
`thereby transforming
`Phase modulation ofan input beam is preferred for creating
`relies on the intensities of
`traps because trapping
`optical
`beams and not on their relative phases. Amplitude modula-
`their
`light away from traps and diminish
`tions may divert
`effectiveness.
`forces
`When a particle is optically trapped, optical gradient
`exerted by the trap exceed other radiation pressures
`arising
`from scattering and absorption. For a Gaussian TEM« input
`laser beam,
`the beam diameter
`this generally means
`that
`the diameter of the
`coincide with
`should
`substantially
`entrance pupil. A preferred minimum numerical
`aperture to
`form a trap is about 0.9 to about 1.0.
`technol-
`One difficulty in implementing
`optical
`trapping
`ogy is that each trap to be generated generally requires its own
`focused beam of light. Many systems of interest
`require mul-
`traps, and several methods have been developed
`tiple optical
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`to achieve multiple trap configurations. One existing method
`is redirected between multiple
`uses a single light beam that
`to "time-share"
`trap locations
`the beam between various
`traps. However, as the number oftraps increases,
`the intervals
`during which each trap is in its "ofl" state may become long
`for particles to diffuse away from the trap location before the
`trap is re-energized. All these concerns have limited imple-
`mentations of this method to less than about 10 traps per
`system.
`traditional method of creating multi-trap
`Another
`systems
`beams of light
`relies on simultaneously
`passing multiple
`aperture lens. This is done by
`through a single high numerical
`lasers or by using one or more beam
`either using multiple
`in the beam of a single laser. One problem with this
`splitters
`technique is that, as the number oftraps increases,
`the optical
`system becomes progressively more and more complex.
`Because of these problems,
`of
`the known implementations
`this method are limited to less than about 5 traps per system.
`In a third approach for achieving a multi-trap
`system, a
`diffractive optical element (DOE) (e.g., a phase shifting holo-
`gram utilizing either a transmission or a reflection geometry)
`is used to alter a single laser beam's wave front. This invention
`is disclosed in U.S. Pat. No. 6,055, 106 to Grier et al. The
`is altered so that the downstream laser beam essen-
`wavefront
`tially becomes a large number of individual
`laser beams with
`and directions of travel
`fixed by the exact
`relative positions
`nature of the diffractive
`optical element. In effect, the Fourier
`transform of the DOE produces a set of intensity peaks each
`trap or "tweezer. "
`of which act as an individual
`of the third approach have used a
`Some implementations
`hologram to create between 16 and 400
`fixed transmission
`trapping centers.
`individual
`A fixed hologram has been used to demonstrate
`the prin-
`ciple of holographic optical trapping but using a liquid crystal
`grating as the hologram permitted 'manufacture' of a separate
`hologram for each new distribution of traps. The spatially
`imposed on the trapping laser by
`varying phase modulation
`the liquid crystal grating may be easily controlled in real time
`a variety of dynamic manipu-
`by a computer,
`thus permitting
`lations.
`types of traps
`that may be used to optically trap
`Other
`particles include, but are not limited to, optical vortices, opti-
`cal bottles, optical rotators and light cages. An optical vortex
`an area of zero electric field
`produces a gradient
`surrounding
`particles with dielectric con-
`to manipulate
`which is useful
`than the surrounding medium or which are
`lower
`stants
`reflective, or other types of particles which are repelled by an
`optical trap. To minimize its energy, such a particle will move
`to the region where the electric field is the lowest, namely the
`zero electric field area at the focal point of an appropriately
`shaped laser beam. The optical vortex provides an area of zero
`(toroid). The
`electric field much like the hole in a doughnut
`is radial with the highest electric field at the
`optical gradient
`circumference of the doughnut. The optical vortex detains a
`small particle within the hole of the doughnut. The detention
`is accomplished by slipping the vortex over the small particle
`along the line of zero electric field.
`The optical bottle differs from an optical vortex in that
`it
`has a zero electric field only at the focus and a non-zero
`electric field in all other directions
`the focus, at
`surrounding
`an end of the vortex. An optical bottle may be useful
`in
`trapping atoms and nanoclusters which may be too small or
`twee-
`too absorptive to trap with an optical vortex or optical
`zers. (See J. Arlt and M. J. Padgett. "Generation of a beam
`by regions of higher
`with a dark focus surrounded
`intensity:
`The optical bottle beam, "Opt. Lett. 25, 191-193,2000.)
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`US 8,933,395 B2
`
`The light cage (U.S. Pat. No. 5,939,716) is loosely, a mac-
`roscopic cousin of the optical vortex. A light cage forms a
`ring of optical
`traps to surround a particle too
`time-averaged
`large or reflective
`to be trapped with dielectric
`constants
`lower than the surrounding medium.
`When the laser beam is directed through or reflected from the
`phase patterning optical element,
`the phase patterning optical
`a plurality of beamlets having an altered
`element produces
`and type of optical
`phase profile. Depending on the number
`the alteration may include diffraction, wave-
`traps desired,
`and con-
`front
`phase shifting,
`steering,
`diverging
`shaping,
`verging. Based upon the phase profile chosen,
`the phase pat-
`terning optical element may be used to generate optical
`traps
`in the form of optical
`traps, optical vortices, optical bottles,
`light cages, and combinations oftwo or more
`optical rotators,
`of these forms.
`for manipulat-
`Researchers have sought
`indirect methods
`the cells with diamond micro-
`ing cells,
`such as tagging
`particles. Cell
`particles
`the diamond
`and then tweezing
`have included cell orientation for microscopic
`manipulations
`analysis as well as stretching cells. Tissue cells have also been
`in vitro in the same spatial distribu-
`arranged with tweezers
`tion as in vivo.
`In addition to the cells themselves,
`tweezers have
`optical
`been used to manipulate
`such as vesicles
`cellular organelles,
`or globular
`chromosomes,
`transported
`along microtubules,
`DNA. Objects have also been inserted into cells using optical
`hveezers.
`as an example of new types of sorting using
`Accordingly,
`traps, a method of cell sorting using a
`laser steered optical
`technique which isolates valuable cells from other cells, tis-
`is needed. Further, a way of achieving
`sues, and contaminants
`a unique contribution of optical
`trapping to the major indus-
`trial needs of blood cell sorting and purification is required.
`there is a need to separate
`sperm cells in the
`Still further,
`animal husbandry market.
`for a separation tech-
`As a consequence,
`a need remains
`nique and apparatus which is continuous, has high through-
`and which causes negligible or
`time saving,
`put, provides
`for separation.
`In
`minimal damage to the various components
`should have further applicability to
`such techniques
`addition,
`biological or medical areas, such as for separations of blood,
`sperm, other cellular materials, as well as viral, cell organelle,
`and other biologi-
`colloidal suspensions,
`structures,
`globular
`cal materials.
`
`SUMMARY OF THE INVENTION
`
`The exemplary embodiments of the present
`invention pro-
`vide for separating components
`in a mixture,
`such as separat-
`ing the various blood components of whole blood into corre-
`sponding fractions, such as a platelet fraction, a red blood cell
`fraction, a white blood cell fraction, and a plasma fraction.
`of the present
`The various
`embodiments
`invention provide
`separation of components
`on a continuous
`such as
`basis,
`closed system, without
`a continuous,
`the potential
`within
`damage and contamination of prior art methods, particularly
`for fractionation of blood components. The continuous pro-
`cess of the present
`time
`also provides
`significant
`invention
`for blood fractionation.
`In
`and higher
`savings
`throughput
`embodiments may also include addi-
`the various
`addition,
`for separating
`the compo-
`tional means
`and manipulating
`and
`optical manipulation
`holographic
`particularly
`nents,
`separation. The various embodiments may also be applied to
`separations of other types ofcellular and biological materials,
`such as sperm, viruses, bacteria, cell debris, cell organelles,
`
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`suspensions,
`
`cellular debris,
`
`colloidal
`structures,
`globular
`and other biological materials.
`As used herein, "Particle" refers to a biological or other
`but not limited to, oligonucle-
`chemical material
`including,
`otides, polynucleotides,
`chemical compounds,
`lip-
`proteins,
`antigens, cel-
`ligands, cells, antibodies,
`ids, polysaccharides,
`of cells,
`lipids, blastomeres,
`lular organelles,
`aggregations
`peptides, cDNA, RNA and the like.
`microorganisms,
`An exempla