J. Inst. Electrostat. Jpn.

, 30, 3 (2006) 140 145⚦ሹ⛘✼⤑ઃ߈⺃㔚ᵒേ࠺ࡃࠗࠬߦࠃࠆ⚦⢩ߩㅪ⛯ಽ㔌▫↰ ఝ 㧘චᨑౝరሶ 㧘⣁ဈ ཅ৻ (2005 ᐕ 8 ᦬ 1 ᣣฃઃ㧧2006 ᐕ 4 ᦬ 25 ᣣฃℂ)

Continuous Separation of Cells Using Dielectrophoretic Device with Porous Insulation
Membrane
Masaru HAKODA*1, Motoko TOSHINAI* and Yoshikazu WAKIZAKA**(Received August 1, 2005; Accepted April 25, 2006)

Dielectrophoresis (DEP) is a motion of dielectric particles caused by polarization effects in a non-uniform
electric field. We apply the DEP to separate the viable and non-viable cells by considering the difference ofpermittivity between viable and non-viable cells. The development of the separator of the viable and the non-viable
cell using DEP device which arranged a porous insulation membrane between the parallel plate electrodes wasexamined. Therefore, DEP velocities of cells were measured under the various conditions of the pore-diameter andthe thickness of the membrane. Furthermore, DEP velocities of cells were analyzed by the simulation and it
compared with the experimental value. Moreover, the separation of the viable and the non-viable cells was carriedout using the continuous DEP separator. And the retention ratio of the viable cell was examined under the various
conditions of flow rate, applied voltage, and electric field frequency. The retention ratio of the viable cell was greaterthan 90 % under the condition of 1 kHz, 70 Vpp, flow rate 0.3 mL/min. It is found that the porous insulation
membrane type DEP separator was effective for the separation of the viable and the non-viable cell.
䋱䋮䈲䈛䉄䈮㩷䈩ታ↪⊛䈭䉴䉬䊷䊦䈱⺃㔚ᵒേ䊐䉞䊦䉺䊷䉕⹜૞䈚䋬䈠䈱᦭⚦⢩䈱ၭ㙃䈮䈲䋬䊋䉟䉥䊥䉝䉪䉺䊷䈏೑↪䈘䉏䈩䈇䉎䈏䋬ၭലᕈ䉕ᬌ⸛䈚䈩䈇䉎 8-11)䋮㩷

㙃ㆊ⒟䈪ᱫ⚦⢩ᢙ䈏Ⴧട䈜䉎䋮ᱫ⚦⢩䈲䋬↢⚦⢩䉇ၭ࿾䈻䈠䈖䈪䋬ᧄ⎇ⓥ䈪䈲⺃㔚ᵒേ䈮䉋䉎ᱫ⚦⢩䈱ㅪ⛯ಽ㔌ⵝᖡᓇ㗀䉕෸䈿䈜น⢻ᕈ䈏䈅䉎䈢䉄䋬䊋䉟䉥䊥䉝䉪䉺䊷ౝ䈱ᱫ⟎䈱㐿⊒䉕⋡ᜰ䈚䋬⚦ሹ⛘✼⤑ဳ⺃㔚ᵒേ䊂䊋䉟䉴䈱᦭ല⚦⢩䉕ㆬᛯ⊛䈮㒰෰䈜䉎䈖䈫䈲ᄢᄌ㊀ⷐ䈪䈅䉎䋮ᣢᓔ䈱⎇ⓥᕈ䉕ᬌ⸛䈜䉎䈢䉄䋬න৻䈱⚦ሹ䈮䈍䈔䉎㔚႐䉕᭽䇱䈭᧦ઙ䈪䉋䉍䋬↢⚦⢩䈫ᱫ⚦⢩䈲䈠䉏䈡䉏⺃㔚₸䈏⇣䈭䉎䈢䉄䋬⺃㔚⸃ᨆ䈚䋬⚦⢩䉕↪䈇䈩⚦ሹㄭற䈱⺃㔚ᵒേㅦᐲ䉕᷹ቯ䈚䈢䋮ᵒേ䉕ⴕ䈉䈫䈠䈱ᵒേ⁁ᘒ䈮㆑䈇䈏⷗䉌䉏䉎䈖䈫䈏ႎ๔䈘䉏䈩䈘䉌䈮䋬ᄙሹᕈ⛘✼⤑䉕↪䈇䈢↢⚦⢩䈫ᱫ⚦⢩䈱ಽ㔌䉕ⴕ䈇䋬1-7)

䋮䈚䈎䈚䋬䈠䉏䉌䈱⎇ⓥ䈲⭯⤑㔚ᭂ䈮䉋䉎䉅䈱䈪䋬䊙䉟

ㅘㆊᶧᵹ㊂䋬ශട㔚࿶䋬๟ᵄᢙ䈭䈬䈱ᠲ૞᧦ઙ䉕ᄌൻ䈘䈞䈩䉪䊨䉼䉾䊒਄䈪䈱ಽ㔌䉕⋡⊛䈮䈚䈢䉅䈱䈪䈅䉍䋬ᄢ㊂ಣℂ䈜䉎↢⚦⢩଻ᜬ₸䈍䉋䈶ᱫ⚦⢩଻ᜬ₸䉕᷹ቯ䈚䋬ᧄⵝ⟎䈮䉋䉎↪ㅜ䈮䈲ะ䈇䈩䈇䈭䈇䋮䈠䈖䈪䋬ᚒ䇱䈲䊪䉟䊟䊷㔚ᭂ䉕↪䈇㩷↢ᱫ⚦⢩ಽ㔌䈱᦭ലᕈ䉕ᬌ⸛䈜䉎䈫䈫䉅䈮ᦨㆡ᧦ઙ䉕᣿䉌䈎䈇䉎䈮䈜䉎䈖䈫䉕⋡⊛䈫䈜䉎䋮㩷ࠠ࡯ࡢ࡯࠼㧦⺃㔚ᵒേ䋬䊐䉞䊦䉺䊷䋬↢ᱫ⚦⢩䋬⚦⢩ಽ㔌䋬㩷ਇဋ৻㔚⇇㩷* ⟲㚍ᄢቇᎿቇㇱ㩷 ↢‛ൻቇᎿቇ⑼㩷 㩿376-8515 ⟲㚍⋵᩿↢Ꮢᄤ␹↸㩷 1-5-1㪀㩷
Department of Biological and Chemical Engineering,Gunma University, Kiryu, Gunma 376-8515, Japan** ᄢ㒋ᄢቇ CASI 㩿ᄢ㒋ᐭ็↰Ꮢጊ↰ਐ 2-1㪀㩷
Center for Advanced Science and Innovation,
Osaka University1
hakoda@bce.gunma-u.ac.jp䋲䋮⺃㔚ᵒേℂ⺰ 12)㩷☸ሶ䉕㔚႐ਛ䈮䈍䈒䈫ಽᭂ䈚䋬ಽᭂ䈪↢䈛䈢ᱜ䈍䉋䈶⽶䈱㔚⩄䈲䋬㔚ᭂ䈱⽶㔚ᭂ䈍䉋䈶ᱜ㔚ᭂ䈫䈏ᒁ䈐䈧䈔䈅䈉䋮ဋ৻䈭㔚႐䈱႐ว䋬ᱜ㔚ᭂ䈫☸ሶ䈱⽶㔚⩄䈏ᒁ䈐ว䈉ജ䈫䋬⽶㔚ᭂ䈫☸ሶ䈱ᱜ㔚⩄䈏ᒁ䈐ว䈉ജ䈲╬䈚䈒䋬㔚⩄䉕ᜬ䈢䈭䈇☸
ሶ䈱㔚᳇ജ䈲⋧Ვ䈘䉏䉎䈢䉄☸ሶ䈲⒖േ䈚䈭䈇䋮Fig.1 䈱䉋䈉䈮䋬ਇဋ৻㔚႐䈱႐ว䋬☸ሶ䈱⺃㔚₸䈏๟࿐ᇦ⾰䈱⺃㔚₸141 (37)䉋䉍ᄢ䈐䈇䈫䈐䋬☸ሶ䈲ᒝ㔚႐஥䈮⒖േ䈜䉎㩿ᱜ䈱⺃㔚ᵒേ㪀䋮䊦䉮䊷䉴ṁᶧ䈮 Ộ䈚䈢䋮䉁䈢䋬☸ሶ䈱⺃㔚₸䈏๟࿐ṁᶧ䈱⺃㔚₸䉋䉍ዊ䈘䈇႐ว䋬☸
ሶ䈲ᒙ㔚႐஥䈻⒖േ䈜䉎㩿⽶䈱⺃㔚ᵒേ㪀䋮㩷㱑m⺃㔚ᵒേജ FDEP 䈲ℂ⺰⊛䈮ᰴ䈱䉋䈉䈮⴫䈘䉏䉎䋮㩷

Hc  Hm
E 2H c *  2H m**FDEP2S r 3H m䈖䈖䈪䋬r: ☸ሶ䈱ඨᓘ䋬İm:

㱑H*㱑LǭH ǭm ǭL(1)ǭOpermittivity of medium

Pin electrode๟࿐ᇦ⾰䈱Ყ⺃㔚₸䋬İm*:๟࿐ᇦ⾰䈱ⶄ⚛⺃㔚₸䋬İ䌣*: ⚦⢩䈱ⶄ⚛⺃㔚₸䋬E: 㔚႐ᒝᐲ䈪

Fig.1 Dielectrophoretic mobility of bio-particles
under non-uniform electric field.

䈅䉎䋮㩷䉴䊃䊷䉪䉴䈱ᛶ᛫ᴺೣ䉕೑↪䈜䉎䈖䈫䈮䉋䉍䋬ℂ⺰⊛䈭⺃㔚ᵒേㅦᐲ䉕᳞䉄䉎䈖䈫䈏䈪䈐䉎䋮☼ᕈ₸䉕㱓䋬⺃㔚ᵒേㅦᐲ䉕ElectrodevDEP 䈫䈜䉎䈫䋬☼ᕈജ Fd 䈲ᰴ䈱䉋䈉䈮⴫䈘䉏䉎䋮㩷Fd6SK rvDEP

Insulation membrane(2)Fd䋽FDEP 䈫઒ቯ䈜䉎䈫䋬⺃㔚ᵒേㅦᐲ vDE㪧 䈲ᰴ䈱䉋䈉䈮⴫䈘Electrode䉏䉎䋮㩷r 2H m H c  H m
E 2

3K H c *  2H m**vDEP

*(3)

Pore

Fig.2 Non-uniform electric field by parallel plateelectrodes and pore membrane.

ੑᨎ䈱᧼⁁㔚ᭂ䈱㑆䈮⚦ሹ⛘✼⤑䉕᜽䉃䈖䈫䈮䉋䈦䈩 Fig.2
䈱䉋䈉䈭ਇဋ৻㔚႐䈏ᒻᚑ䈘䉏䋬⛘✼⤑䉋䉍ᄢ䈐䈭⺃㔚₸䉕

Monitor

CCD-cameraᜬ䈧ṁᶧ䉕↪䈇䈢႐ว䈮䈲䋬⚦ሹㄭற䈏ᒝ㔚႐䈮䈭䉍䋬㔚ᭂㄭற䈏ᒙ㔚႐䈮䈭䉎䋮㩷

Microscope䋳䋮 ታ㛎ⵝ⟎෸䈶ᣇᴺ㩷
Fig.3 䈮⺃㔚ᵒേㅦᐲ᷹ቯ䈱䈢䉄䈱ⵝ⟎䈱ో૕࿑䉕䋬Fig.4VTR
Dielectrophoretic

Function䈮⺃㔚ᵒേㅦᐲ᷹ቯ↪䊂䊋䉟䉴䈱᭎⇛࿑䉕␜䈜䋮䉝䉪䊥䊦᧼

devicegenerator䋳䋮䋱㩷 න৻⚦ሹ⛘✼⤑ઃ䈐⺃㔚ᵒേ䉶䊦㩷䈪㔚ᭂ㑆〒㔌 㪈mm,㩷 㜞䈘 3 mm, 㐳䈘 30 mm 䈱ᵹ〝䉕૞䉍䋬ੑᨎ䈱䊒䊧䊷䊃㔚ᭂ䈱㑆䈮⚦ሹ⛘✼⤑䉕᜽䉃䈖䈫䈮䉋䉍ਇဋFig.3 Schematic diagram of experimental apparatus ofdielectrophoresis.

৻㔚႐䉕ᒻᚑ䈚䈢䋮䊒䊧䊷䊃㔚ᭂ䈲 3 mm㬍10 mm, ෘ䈘 0.2mm 䈱䊆䉾䉬䊦᧼䉕૶↪䈚䋬⛘✼⤑䈲 3 mm㬍30 mm㪃㩷 ෘ䈘0.2 mm 䈱䊘䊥䉣䉼䊧䊮⤑䉕૶↪䈚䈢䋮䉁䈢㔚ᭂ㑆〒㔌䈲 1Insulationmm䋬⛘✼⤑䈫㔚ᭂ䈱〒㔌䈲 0.4 mm 䈫䈚䈢䋮Fig.3 䈮⺃㔚ᵒേ

membrane㨪ElectrodeFunctionታ㛎ⵝ⟎䈱᭎⇛࿑䉕␜䈜䋮⺃㔚ᵒേ䊂䊋䉟䉴䈮੤ᵹ㔚Ḯ

generator(33250A, Agilent, USA)䉕૶↪䈚䋬੤ᵹ㔚႐䉕ශട䈚䈢䋮䈠䈱䈫䈐䈱⺃㔚ᵒേ䊂䊋䉟䉴ౝ䈱᭽ሶ䉕㗼ᓸ㏜(BHC, OLIMPASCo, Japan)䈮ធ⛯䈘䉏䈢 CCD 䉦䊜䊤(WV-CD20, National Co,Japan)䉕ㅢ䈚䋬䊝䊆䉺䊷(WV-5410, National Co, Japan)䈪⏕⹺䈚䋬න૏ᤨ㑆䈅䈢䉍䈱ᵒേ〒㔌䉋䉍⺃㔚ᵒേㅦᐲ䉕᷹ቯ䈚䈢䋮Viable cell䈠䈱㓙䋬⚦ሹ䈱౉ญㇱ䈮㗼ᓸ㏜䈱䊐䉤䊷䉦䉴䉕ว䉒䈞䈩౉ญ

Non-viable cellઃㄭ䈱⚦ሹਛᔃ✢਄䈱⚦⢩䈱ᵒേㅦᐲ䉕᷹ቯ䈚䈢䋮ታ㛎䈮䈲䊙䉡䉴䊊䉟䊑䊥䊄䊷䊙䈪䈅䉎ᶋㆆᕈ⚦⢩䋬3䋭2H3 ⚦⢩䋨RCB0867, ℂ⎇䉳䊷䊮䊋䊮䉪䋩䈱↢ᱫ⚦⢩䉕૶↪䈚䋬0.3M 䉫Fig.4

Device ofmembrane.

dielectrophoresis

withinsulation

142 (38)

30

32006

䋳䋮䋲㩷 㺂㪜㩷㪉 䈱⸃ᨆ㩷㩷 ੑᨎ䈱ᐔⴕᐔ᧼㔚ᭂ㑆䈮⚦ሹ⛘✼᧼䉕᜽䉃䈖䈫䈮䉋䉍 Fig.2

Cellsuspension

Retention flow䈱䉋䈉䈭ਇဋ৻㔚႐䈏ᒻᚑ䈘䉏䉎䈫⠨䈋䉌䉏䉎䋮䈠䈖䈪⚦ሹ䈱ሹᓘ䉇ᢙ䋬⛘✼⤑䈱ෘ䈘䉕ᄌൻ䈘䈞䈢䈫䈐䈱㺂E2 䈱ᄌൻ䉕᦭Pump㒢ⷐ⚛ᴺ䈮䉋䈦䈩⸃ᨆ䈚䈢䋮᦭㒢ⷐ⚛ᴺ䈮䉋䈦䈩᳞䉄䉌䉏䈢㔚૏䈎䉌㔚႐䉕᳞䉄䋬㔚႐䈎䉌㺂E2 䉕᳞䉄䈢䋮⸃ᨆ䈮䈲 㪉 ᰴ

DEP separator

Function
generatorర᦭㒢ⷐ⚛ᴺ⸃ᨆ䉸䊐䊃(Maxwell 2D, Ansoft Corp., USA)䉕
૶↪䈚䈢䋮㩷
Pump㩷
䋳䋮䋳㩷 ⚦ሹ⛘✼⤑ઃ䈐⺃㔚ᵒേ䊂䊋䉟䉴㩷Fig.5⚦ሹ⛘✼⤑ઃ䈐⺃㔚ᵒേ䊂䊋䉟䉴䉕↪䈇䈢↢ᱫ⚦⢩䈱ㅪ

Schematic

diagramofcontinuous

dielectrophoretic separation apparatus.

⛯ᑼಽ㔌䈮䈍䈔䉎ታ㛎ⵝ⟎䈱ో૕࿑䉕 Fig.5 䈮␜䈜䋮⚦⢩ Ộᶧ䈲䊘䊮䊒䈮䉋䉍⺃㔚ᵒേ䊂䊋䉟䉴䈮ଏ⛎䈘䉏䋬଻ᜬᶧ஥

Acrylic resin䈫ㅘㆊᶧ஥䈮ಽ䈎䉏䈩ឃ಴䈘䉏䉎䋮⚦⢩ Ộᶧ䈫ㅘㆊᶧ஥䈱Ni electrodeᵹ㊂䈲䊘䊮䊒䈮䉋䉍⸳ቯ䈚䈢䋮⚦ሹ⛘✼⤑ઃ䈐ㅪ⛯ಽ㔌ⵝ⟎

Silicon plate䈱౮⌀䈫ౝㇱ䈱࿑䉕 Fig.6 䈮␜䈜䋮㔚ᭂ䈱᧚⾰䈲䊆䉾䉬䊦䋬ᄢ
30

䈐䈘䈲 3cm㬍10cm㬍0.3mm 䈫䈚䋬⛘✼⤑䈱᧚⾰䈲䊘䊥䉣䉼䊧䊮䋬ᄢ䈐䈘䈲 3cm㬍8.6cm㬍0.2mm 䈫䈚䈢䋮ෘ䈘 1mm 䈱䉲䊥䉮

Porous86䊮᧼䉕䈒䉍ᛮ䈐 3mm㬍30cm 䈱ᵹ〝ㇱ䉕૞ᚑ䈚䈢䇯䉁䈢䋬⛘✼insulation

membrane⤑䈮䈲⚂ 600㱘m 䈱ሹ㑆〒㔌䈪ሹᓘ 200㱘m㱢䈱⚦ሹ䈏
1000 ୘䈅䉎䋮⚦⢩䈲 3-2H3 ⚦⢩䉕૶↪䈚䋬8.5wt%䉴䉪䊨䊷䉴
䋫0.3wt%䉫䊦䉮䊷䉴ṁᶧ䈮 Ộ䈚䈢䋮⚂ 106cells/mL 䈱⚦⢩ Ộᶧ䉕ᵈ౉䈚䋬଻ᜬᶧ஥䈫ㅘㆊᶧ஥䈎䉌䈠䉏䈡䉏 Ộᶧ䉕

Pore(200㱘mǾ)࿁෼䈚䋬ⴊ⃿⸘▚⋚䈮䉋䉍↢⚦⢩ᢙ䈫ᱫ⚦⢩ᢙ䉕⸘ᢙ䈚䈢䋮଻ᜬ₸䈲䋬଻ᜬᶧ஥ xA 䈫ㅘㆊᶧ஥ xB 䈱↢⚦⢩䈫ᱫ⚦⢩䈠䉏
1 mmEnlarged photo䈡䉏䈱⚦⢩ኒᐲ䈪䈅䉎䋮㩷xA
଻ᜬ₸[㧑]㧩
u 100x A  xB

DEP device photo(1)Fig.6 DEP device with porous insulation membrane.

㩷䋴䋮㩷 ታ㛎⚿ᨐ䈍䉋䈶⠨ኤ㩷

70䋴䋮䋱㩷 න৻⚦ሹ⛘✼⤑ઃ䈐⺃㔚ᵒേ䉶䊦㩷ᐲ䈱᷹ቯ䉕ⴕ䈦䈢䋮න৻⚦ሹ䈎䉌䈱〒㔌䈫⺃㔚ᵒേㅦᐲ䈱㑐ଥ䉕 Fig.7 䈮␜䈜䋮䈖䈖䈪䋬↪䈇䈢⚦⢩䈲↢⚦⢩䈱䉂䈮䈧䈇䈩ᬌ⸛䈜䉎䋮ᱫ⚦⢩䈲⺃㔚ᵒേജ䉕ᱴ䉖䈬ฃ䈔䈭䈇䈢䉄⺃㔚ᵒേ䈲↢䈛䈭䈇䋮⺃㔚ᵒേㅦᐲ䈲⚦ሹ䈮ㄭ䈨䈒䈮ᓥ䈇Ⴧᄢ䈚䈢䋮䉁䈢䋬㔚࿶䈏㜞䈇䈾䈬⺃㔚ᵒേㅦᐲ䈏ㅦ䈒䈭䉎䈫䈇䈉⚿ᨐ䈏ᓧ䉌䉏䈢䋮Fig.8 䈮⛘✼⤑䈱ෘ䈘 0.3mm 䈫 0.5mm 䈱႐ว䈮䈍䈔䉎ሹᓘ䈫⺃㔚ᵒേㅦᐲ䈱㑐ଥ䈮䈧䈇䈩䈱ታ㛎⚿ᨐ䉕␜䈜䋮⺃㔚ᵒേㅦᐲ䉕Ყセ䈚䈢૏⟎䈲න৻⚦ሹ䈱ਛᔃ✢਄䈪⛘✼⤑䈎䉌 0.1mm 䈱䈫䈖䉐䈱ᵒേㅦᐲ䉕ᬌ⸛䈚䈢䋮Ყセ䈚䈢

DEP velocity[µm/s]න৻⚦ሹ⛘✼⤑䉕↪䈇䈩䋬⚦ሹㄭற䈮䈍䈔䉎⺃㔚ᵒേㅦ

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300

400

Distance from membrane [Ǵm]⛘✼⤑䈱ෘ䈘 0.3mm 䈱ᣇ䈏 0.5mm 䉋䉍䉅⺃㔚ᵒേㅦᐲ䈏ᄢ䈐䈇䈫䈇䈉⚿ᨐ䈏ᓧ䉌䉏䋬⛘✼⤑䈱ෘ䈘 0.3mm 䈱䈫䈐䋬ሹᓘFig.7 Correlation between dielectrophoretic velocity and300㱘m 䈱⺃㔚ᵒേㅦᐲ䈏ᦨᄢ䈮䈭䉍䋬⛘✼⤑䈱ෘ䈘 0.5mm

distance from membrane (Pore diameter: 200㱘m,䈱䈫䈐䋬ሹᓘ 400㱘m 䈱ᵒേㅦᐲ䈏ᦨᄢ䈮䈭䈦䈢䋮㩷

Thickness: 3mm, Frequency: 1MHz)).

143 (39)ᓟㅀ䈜䉎⸃ᨆ⚿ᨐ䈪䈲㺂E2 䈲ሹᓘ䈏ዊ䈘䈇ᣇ䈏ᄢ䈐䈇䈏䋬

Pore center⺃㔚ᵒേㅦᐲ䈲․ቯ䈱ሹᓘ䈪ᦨᄢ୯䉕␜䈚䈢䋮䈖䈱ේ࿃䈲䋬Electrodeᶧ䈱ᵹ䉏䈏䈭䈇䈢䉄䋬ሹᓘ䈏ዊ䈘䈇႐ว䋬⚦ሹౝ䈻⚦⢩䈏䈧䉁䉍⚦ሹ䈱ᵒേ䈏ᛥ೙䈘䉏䉎䈢䉄䈫⠨䈋䉌䉏䉎䋮㩷
ШE 2 [V2/m3]
2䋴䋮䋲㩷 㺂E 䈱⸃ᨆ㩷⚦⢩ಽ㔌䈱น⢻ᕈ䉕ᬌ⸛䈜䉎䈢䉄䋬⚦ሹㄭற䈱㺂E2 䈱䉲䊚Insulation membrane䊠䊧䊷䉲䊢䊮䉕ⴕ䈦䈢䋮㺂E2 䈱䉲䊚䊠䊧䊷䉲䊢䊮䈲䋬⚦ሹᓘ䋬⛘✼⤑䈱ෘ䈘䋬⚦ሹ㑆〒㔌䉕ᄌൻ䈘䈞䋬᦭㒢ⷐ⚛ᴺ䈮䉋䉍ⴕ䈦
䈢䋮Fig.9 䈲䋬਄ਅ䈱㔚ᭂ䈮᜽䉁䉏䈢⚦ሹ䈱䈅䉎⛘✼⤑䈮䈍䈔
䉎㺂E2 䈱ಽᏓ࿑䉕␜䈜䋮䈖䈱⸃ᨆ䈲౞╴ᐳᮡ䈪ⴕ䈇䋬ਛᔃ✢஥䉕ឬ䈇䈩䈇䉎䋮⊕䈇䈾䈬㺂E2 䈏ᄢ䈐䈒䋬㤥䈒䈭䉎䈾䈬䈏䈱

㺂E2 ዊ䈘䈇䈖䈫䉕␜䈚䈩䈇䉎䋮䈠䈱⚿ᨐ䋬㺂E2 䈲⛘✼⤑䈱⚦Electrodeሹ䈱ⷺㄭற䈏ᄢ䈐䈒䈭䉎䈖䈫䈏䉒䈎䉎䋮䉁䈢䋬⚦ሹ䈱ਛᔃゲᣇ

Fig.9 Distribution of ШE2 between electrodes.

ะ䈪䈲䋬⚦ሹ䈱౉ญઃㄭ䈱㺂E2 䈏ᦨ䉅ᄢ䈐䈒䈭䉎䈖䈫䈏䉒䈎䈦(×1010)
60

䈢䋮㩷
Fig.10 䈲㺂E2 䈮෸䈿䈜ሹᓘ䈱ᓇ㗀䉕␜䈚䈢䋮ሹᓘ 100, 200,50100㱘m䈲ੑ䈧䈱㔚ᭂ㑆䈱૏⟎䉕␜䈚䈩䈇䉎䋮⚿ᨐ䉋䉍䋬ሹᓘ䈏ዊ䈘䈇

40200㱘mⴕ䈉႐ว䋬⚦ሹㄭற䈪䈱෻⊒ജ䈏㊀ⷐ䈫䈭䉎䈢䉄䋬ሹᓘ䈫⚦
2ሹਛᔃㇱ䈱㺂E 䈱㑐ଥ䉕 Fig.11 䈮␜䈜䋮⚦ሹ౉ญ䈮䈍䈔䉎⚦ሹਛᔃㇱ䈪䈲ሹᓘ䈏ዊ䈘䈇䈾䈬㺂E2 䈱୯䈏ᄢ䈐䈒䈭䉍䋬ሹ
ᓘ 200㱘m એਅ䈪ᕆỗ䈮㺂E2 䈏ᄢ䈐䈒䈭䉎䈖䈫䈏䉒䈎䈦䈢䋮㩷

3230

2

䈾䈬㺂E2 䈱୯䈏ᄢ䈐䈒䈭䉎䈖䈫䈏䉒䈎䈦䈢䋮䉁䈢䋬ㅪ⛯ಽ㔌䉕

㺂E [V /m ]300, 400, 500㱘m 䈮䈍䈔䉎⚦ሹ䈱ਛᔃ䈱㺂E2 䉕᳞䉄䈢䋮ᮮゲ300㱘m

20400㱘m

10500㱘m

0700

0Thickness

500

400

600

800

1000

Distance from electrode [㱘m]Fig.10 Effect of pore diameter on 㺂E2between electrodes.

㪊㪇㪇㱘㫄㪌㪇㪇㱘㫄

400300(×10 9 )2002001001500200300

400

Pore diameter[Ǵm]Fig.8 Effect of pore diameter on DEP velocity.

500

㺂E2 [V2 /m 3 ]DEP velocity[µm/sec]

600

200

100500100200300
400
Pore diameter [㱘m]Fig.11 Effect of pore diameter on ШE2.
(Thickness: 200㱘m)

500

144 (40)3032006ᰴ䈮䋬⚦⢩ಽ㔌䈮෸䈿䈜๟ᵄᢙ䈱ᓇ㗀䈮䈧䈇䈩ᬌ⸛䈚䈢䋮(×10 9 )ታ㛎᧦ઙ䈲㔚࿶ 80 Vpp䋬 Ộᶧᵈ౉ᵹ㊂ 1 mL/min䋬ㅘㆊᶧ

200஥䈱ᵹ㊂䉕 0.3 mL/min 䈫䈚䈢䋮๟ᵄᢙ䈲 1, 10, 100, 1 k䋬10 k䋬
50 kHz 䈫ᄌൻ䈘䈞䈢䋮Fig.14 䈲⚦⢩଻ᜬ₸䈮෸䈿䈜๟ᵄᢙ

㺂E2 [V2 /m 3 ]150䈱ᓇ㗀䉕␜䈜䋮䈠䈱⚿ᨐ䋬๟ᵄᢙ䈱Ⴧᄢ䈮઻䈦䈩䋬↢⚦⢩଻ᜬ₸䉅Ⴧᄢ䈚䋬1 kHz 䈱䈫䈐↢⚦⢩଻ᜬ₸䈏ᦨᄢ䈫䈭䈦䈢䋮ᱫ100⚦⢩଻ᜬ₸䈲 50%೨ᓟ䉕␜䈚䈢䋮㩷ᦝ䈮䋬⤑ㅘㆊ䈱ᵹ㊂䈮䈧䈇䈩ᬌ⸛䈚䈢䋮ታ㛎᧦ઙ䈲䋬ශട50

㔚࿶ 60 Vpp 䈫 80 Vpp䋬๟ᵄᢙ 1 kHz 䈪䈅䉍䋬 Ộᶧᵈ౉ᵹ㊂䈲 1 mL/min 䈪৻ቯ䈫䈚䋬ㅘㆊᶧ஥䈱ᵹ㊂䉕 0.2, 0.35, 0.5,0100

200

300

400

500

0.65 mL/min 䈫ᄌൻ䈘䈞䈢䋮Fig.15 䈲⚦⢩଻ᜬ₸䈮෸䈿䈜ᵹ㊂䈱ᓇ㗀䉕␜䈜䋮䈠䈱⚿ᨐ䋬ශട㔚࿶ 60 Vpp 䈪䈲䋬ㅘㆊᶧ஥T hickness of membrane[ 㱘m]䈱ᵹ㊂䉕ዊ䈘䈒䈜䉎䈾䈬↢⚦⢩଻ᜬ₸䉅ᱫ⚦⢩଻ᜬ₸䉅ᄢ䈐
Fig.12 Effect of membrane thickness on ШE2.

䈒䈭䈦䈢䈏䋬80 Vpp 䈪䈲䋬䈾䈿৻ቯ䈪䈅䈦䈢䋮㩷(Pore diameter: 200㱘m)ᰴ䈮䋬䉋䉍ᄙ䈒䈱ᱫ⚦⢩䉕㒰෰䈜䉎䈢䉄䈮ㅘㆊᶧ஥䈱ᵹ㊂
䉕 0.5 mL/min䋬 Ộᶧᵈ౉ᵹ㊂䉕 0.7 mL/min 䈮䈚䋬଻ᜬᶧᰴ䈮䋬⛘✼⤑䈱ෘ䈘 100, 200, 300, 400, 500㱘m 䈮䈍䈔䉎⚦஥䈱ᵹ㊂䉕ૐ䈒䈚䈢䋮ታ㛎᧦ઙ䈲䋬㔚࿶ 80 Vpp䋬๟ᵄᢙ 1, 5,ሹ౉ญ䈮䈍䈔䉎ਛᔃ✢਄䈱㺂E2 䉕᳞䉄䈢䋮⛘✼⤑䈱ෘ䈘䈫⚦

10 kHz 䈪ⴕ䈦䈢䋮Fig.16 䈲䋬଻ᜬᶧ஥ᵹ㊂䉕ૐ䈒䈚䈢႐ว䈱2ሹㄭற䈮䈍䈔䉎㺂E 䈱㑐ଥ䉕 Fig.12 䈮␜䈜䋮䈠䈱⚿ᨐ䋬⛘✼
2⤑䈱ෘ䈘䈏⭯䈇䈾䈬㺂E 䈱୯䈏ᄢ䈐䈒䈭䉎䈖䈫䈏䉒䈎䈦䈢䋮㩷
2⚦⢩଻ᜬ₸䈮෸䈿䈜๟ᵄᢙ䈱ᓇ㗀䉕␜䈜䋮䈠䈱⚿ᨐ䋬๟ᵄ
ᢙ 1 kHz 䈪䈲䋬ᱫ⚦⢩଻ᜬ₸䉕㩷 25 %એਅ䈮䈜䉎䈖䈫䈏䈪䈐䈢䋮㩷 ᦝ䈮䋬㺂E 䈱୯䈮෸䈿䈜㔚ᭂ㑆䈮䈍䈔䉎⛘✼⤑䈱૏⟎䋬⚦䈖䈱䉋䈉䈭⚿ᨐ䈮䈭䈦䈢ේ࿃䈫䈚䈩䋬ᱫ⚦⢩䈲↢⚦⢩䈱⽶䈱⺃ሹ㑆䈱〒㔌䈱ᓇ㗀䈮䈧䈇䈩ᬌ⸛䉕ⴕ䈦䈢䋮䈠䈱⚿ᨐ䋬⛘✼⤑㔚ᵒേജ䈮Ყ䈼䈩ዊ䈘䈇䈏ᱜ䈱⺃㔚ᵒേജ䈏௛䈇䈩䈇䉎䈢䈱૏⟎䈲ੑᨎ䈱㔚ᭂ䈱ਛᔃ䈏ᦨㆡ䈪䈅䈦䈢䋮⚦ሹ㑆䈱〒㔌䉄䋬଻ᜬᶧ஥䈱ᵹ㊂䉕ዊ䈘䈒䈜䉎䈖䈫䈮䉋䈦䈩ᱫ⚦⢩䉕⚦ሹ䈻

2

䈲䋬〒㔌䈏⁜䈒䈭䉎䈾䈬㺂E 䈲ዊ䈘䈒䈭䉎䈏䋬400㱘m એ਄䈪ᒁ䈐ነ䈞䉎䈖䈫䈏䈪䈐䈢䈱䈪䈲䈭䈇䈎䈫⠨䈋䉌䉏䉎䋮㩷䈲䈾䈫䉖䈬ᄌ䉒䉌䈭䈇䈫䈇䈉䈖䈫䈏䉒䈎䈦䈢䋮㩷100䋴䋮䋳㩷 ⚦ሹ⛘✼⤑ઃ⺃㔚ᵒേ䊂䊋䉟䉴䈮䉋䉎⚦⢩ಽ㔌㩷䈱᷹ቯ⚿ᨐ䉕䉅䈫䈮⚦ሹ⛘✼⤑ઃ䈐⺃㔚ᵒേ䊂䊋䉟䉴䉕↪䈇䈢↢ᱫ⚦⢩䈱ㅪ⛯ಽ㔌䉕ⴕ䈦䈢䋮ታ㛎᧦ઙ䈲๟ᵄᢙ 5 kHz䋬 Ộᶧᵈ౉ᵹ㊂ 1 mL/min䋬ㅘㆊᶧ஥䈱ᵹ㊂䉕 0.3 mL/min 䈫䈚䈢䋮㔚࿶䈲 40, 50, 60, 70, 80Vpp 䈫ᄌൻ䈘䈞䈢䋮⚦⢩଻ᜬ₸䈮෸䈿䈜㔚࿶䈱ᓇ㗀䉕 Fig.13 䈮␜䈜䋮䈠䈱⚿ᨐ䋬

80
Retention ratio[䋦]න৻⚦ሹ⛘✼⤑䈮䈍䈔䉎㺂E2 䈱⸃ᨆ⚿ᨐ䈫⺃㔚ᵒേㅦᐲViable cell

Theoretical value 53.6Vpp

60
40

Non-viable cell20㔚࿶䉕㜞䈒䈜䉎䈾䈬↢⚦⢩଻ᜬ₸䈲ᄢ䈐䈒䈭䉍䋬ᱫ⚦⢩଻ᜬ₸䈲 50䋦೨ᓟ䈪䈅䈦䈢䋮䈖䈱⚿ᨐ䉋䉍䋬↢⚦⢩䈲 60Vpp એ਄䈪䈲 90%⒟ᐲ䈱଻ᜬ₸䈏ᓧ䉌䉏䈢䋮䉁䈢䋬⤑䈱ㅘㆊᵹㅦ䈫⺃㔚ᵒേㅦᐲ䈏╬䈚䈇႐ว䋬⚦⢩䈲⤑䉕ㅘㆊ䈚䈭䈒䈭䉎䋮䈠䈱䈫䈐䈱ශട㔚࿶䉕ℂ⺰ශട㔚࿶䈫ቯ⟵䈚䋬ℂ⺰ශട㔚࿶䉕▚಴䈚䈢䋮ℂ⺰ශട㔚࿶䉕᳞䉄䉎䈢䉄䈮ᔅⷐ䈭⺃㔚ᵒേㅦᐲ䈫㺂E2 䈱㑐ଥ䈲䋬䊏䊮㪄䊪䉟䊟䊷㔚ᭂ䉕↪䈇䈩ታ᷹䈚䋬ᰴᑼ䉕ᓧ䈢䋮㩷

v DEP  4 K 10 17 b \E 2(4)䈠䈱⚿ᨐ䋬Fig.13 䈱ታ㛎᧦ઙ䈮䈍䈔䉎ℂ⺰ශട㔚࿶ 53.6 Vpp䈪䈅䉍䋬ታ㛎⚿ᨐ䈫⦟䈒৻⥌䈚䈢䋮㩷

040

5060
70
Applied voltage[Vpp]80

Fig.13 Effect of applied voltage on cell retention ratio.

145 (41)䋵䋮䉁䈫䉄㩷
100ᧄ⎇ⓥߪ㧘⚦⢩↪ࡃࠗࠝ࡝ࠕࠢ࠲࡯ౝߩᱫ⚦⢩ࠍㆬᛯ⊛

Retention ratio[䋦]ߦ㒰෰ߢ߈ࠆታ↪⊛ߥಽ㔌ⵝ⟎ߩ㐿⊒ࠍ⋡⊛ߣߒߡⴕࠊ

Viable cell80ࠇߚ㧚ᐔⴕᐔ᧼㔚ᭂ㑆ߦ⚦ሹ⛘✼⤑ࠍ⸳⟎ߔࠆߎߣߦࠃࠅ㧘ਇ

60ဋ৻㔚႐ࠍᒻᚑߒ㧘⚦⢩ࠍ⺃㔚ᵒേߢ߈ࠆߎߣ߇᣿ࠄ߆ߦߥߞߚ㧚߹ߚ㧘㔚႐ߩ⸃ᨆࠃࠅ㧘⚦ሹߩሹᓘ߇ዊߐ޿߶ߤ㧘40߹ߚ⛘✼⤑ߩෘߐ߇⭯޿߶ߤШE2 ߇ᄢ߈ߊߥࠆߣ޿߁⚿Non-viable cellᨐ߇ᓧࠄࠇߚ㧚

20 ߹ߚ㧘⚦ሹ⛘✼⤑ઃ߈⺃㔚ᵒേ࠺ࡃࠗࠬࠍ↪޿ࠆߎߣߦ
ࠃߞߡ㧘↢⚦⢩ߣᱫ⚦⢩ߩಽ㔌߇น⢻ߢ޽ࠆߎߣ߇᣿ࠄ߆

01101001000Frequency[Hz]10000 100000

ߦߥߞߚ㧚ᦝߦ㧘⤑ㅘㆊᵹ㊂㧘ශട㔚࿶㧘๟ᵄᢙߥߤߩᠲ
૞᧦ઙࠍ೙ᓮߔࠆߎߣߦࠃߞߡ㧘90 %એ਄ߩ↢⚦⢩଻ᜬ₸ࠍᓧࠆߎߣ߇ߢ߈ߚ㧚એ਄ߩ⚿ᨐࠃࠅ㧘⚦ሹ⛘✼⤑ઃ߈⺃㔚ᵒേ࠺ࡃࠗࠬߪታ

Fig.14 Effect of frequency on cell retention ratio.

↪⊛ߥ↢ᱫ⚦⢩ߩಽ㔌ⵝ⟎ߢ޽ࠆߎߣ߇␜ໂߐࠇߚ㧚100ෳ⠨ᢥ₂㩷

Retention ratio [%]80

Viable cell: 60Vpp
Non-viable cell: 60Vpp
Viable cell: 80Vpp
Non-viable cell: 80Vpp

601) J.S. Crane, H.A. Pohl: J. Electrochem. Soc., 115, 6, (1968)1752) B.D. Manson, P.M. Townsley: Can. J. Microbiol., 17, (1970)
879403) H.A. Pohl, K. Kaler: Cell Biophys. 1, (1979) 154) G.H. Markx, M.S. Talary, R. Pethig: J. Biotechnol. 32,20(1994) 29
00.2

0.3

0.4

0.5

0.6

0.75) G.H. Markx, R. Pethig: Biotechnol. Bioeng., 45, 4, (1995)
337

㪧㪼㫉㫄㪼㪸㫋㫀㫆㫅㩷㪽㫃㫆㫎㩷㪲㱘㪣㪆㫄㫀㫅㪴6) A. Docosilis, N. Kalogerakis, L.A. Behie, K.V.I.S. Kaler:
Biotechnol. Bioeng., 54, 3, (1997) 239

Fig.15 Effect of permeation flow on cell retention ratio.
7)

100

A.

Docosilis,N.

Kalogerakis,L.A.

Behie:

Cytotechnolology, 30, (1999) 133

Viable cellRetetion ratio [䋦]808) ▫↰ ఝ⣁ဈཅ৻⊕␹⋥ᒄ㧦ൻቇᎿቇ⺰ᢥ㓸, 27(2),(2001) 1599) Y. Wakizaka, M. Hakoda, N. Shiragami: Biochem. Eng. J.,

6020(1), (2004) 1310) Y. Wakizaka, M. Hakoda, N. Shiragami: J. Chem. Eng.

40Japan, 37(7), (2004) 9082011) ▫↰ ఝ⣁ဈཅ৻ਃ੗ା৻ ർፉା⟵㕒㔚᳇

Non-viable cellቇળ⹹䋬29 (2005) 812) H.A. Pohl: Dielectrophoresis, Cambridge University Press

00.1

110100Frequency[kHz]Fig.16 Effect of frequency on retention ratio.
(Permeate flow rate: 0.5mL/min, Feed flow rate:0.7mL/min)(1978)