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- いとは かやぬま
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4 A Light source Power supply B Arbitrary unit (au) (Relative intensity) Arbitrary unit (au) (Relative intensity) Wave length : : nm nm ~20 cm cm Fan Fan3737 o C o C Water filter filter Cultured cells cells Lift Lift C Exposing Wave Length (nm) (nm) TheraBeam VR630: (USHIO LIGHTING, INC.) Light source : : Na-Li lamp Wave length : : 530~630~700 nm nm Power output : : 55 cm: cm: 65mW/cm cm: cm: mw/cm cm: cm: mw/cm cm: cm: mw/cm cm: cm: mw/cm 2 2 Power input : : W 1~15 24 h 3 h min 1 ~ 24 h FACScan analysis Culture in the presence of FBS Culture with 5-ALA in the absence of FBS Culture in the presence of FBS Light exposure Fig.2
5 A. Excitation spectra of PpIX B. Emission spectra of PpIX µm PpIX + Cell 50 µm PpIX + Cell Relative fluorescence intensity Relative fluorescence intensity Excitation Wave Length (nm) Excitation Wave Length (nm) Relative fluorescence intensity Emission Wave Length (nm) C. Excitation spectra of PpIX D. Emission spectra of PpIX U937 (1 mm ALA, 6 h) U937 (1 mm ALA, 6 h) Relative fluorescence intensity Emission Wave Length (nm)
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8 A Scattergram 0 min 1000 Intact FSC Irradiation time 3 min 5 min 10 min 15 min SSC SSC SSC SSC SSC C Propidium iodide stained Irradiation time 0 min 3 min 5 min 10 min 15 min 200 0Counts Damaged FL2-H FL2-H FL2-H FL2-H PI positive FL2-H B Damaged cells (FSC, % of gated) PI positive cells (%) mm ALA 0.25 mm ALA Irradiation time (min) 1 mm ALA 0.25 mm ALA Irradiation time (min) Fig.7 D 15
9 A. Scattergram 0 mm B. Annexin and Propidium iodide stained P I (FL-2H) 1000 FSC UL LL SSC 0 mm UR LR 0.1 mm 0.1 mm 0.25 mm SSC SSC 0.25 mm 0.5 mm SSC 0.5 mm Annexin-V-FITC (FL-1H) Damaged cells (%, FSC) Quadrant (%) ALA (mm) 0 mm 0.1 mm 0.25 mm 0.5 mm UL UR LL LR Fig.8 A. Scattergram 0 min 1000 FSC PI (FL-2H) SSC 5 min SSC 10 min B. Annexin and Propidium iodide stained SSC 15 min SSC 0 min 5 min 10 min 15 min Annexin-V-FITC (FL-1H) Damaged cells (%, FSC) Quadrant (%) Irradiation time (min) 0 min 5 min 10 min 15 min UL UR LL LR Fig.9
10 PI (FL-2H) A. Scattergram 1000 FSC N. I. 0 h 1 h 3 h 6 h 12 h SSC SSC SSC SSC SSC SSC Time after PDT (h) B. Annexin and Propidium iodide stained N. I. 0 h 1 h 3 h 6 h 12 h Annexin-V-FITC (FL-1H) Damaged cells (%, FSC) UL UR LL LR Fig.10 Quadrant (%) 0h 1h 3h 6h 12h
11 1000 FSC A. Scattergram 0 B. TMRE stained 1000 N. I SSC N. I. 3h SSC 3h 12h SSC 12h Damaged cells (FCS, %) C.Time dependent changes Damaged cells Membrane potential Incubation time (h) Decrease of membrane potential (FL-2, % of gated) Count TMRE (FL-2H) Fig.11
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2 1,384,000 2,000,000 1,296,211 1,793,925 38,000 54,500 27,804 43,187 41,000 60,000 31,776 49,017 8,781 18,663 25,000 35,300 3 4 5 6 1,296,211 1,793,925 27,804 43,187 1,275,648 1,753,306 29,387 43,025
USHIO DISCHARGE LAMPS UV LAMPS Deep UV LAMPS XENON LAMPS XENON FLASH LAMPS LAMP HOUSINGS OPTICAL MEASURING INSTRUMENTS USHIO DISCHARGE LAMPS p.4-9 p.10-11 p.1-18 p.-1 p.- 4 p.6-9 7 8 9 p.30-37 UV LAMPS
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E-mail: khatano@fms.saitama-u.ac.jp Tel & Fax: 048-858-3535 Toxin Virus Bacterium Glycolipid Glycoprotein Vero toxin (Stx1 and Stx2) Side view Bottom view H H H H N H Grobotriaosyl Ceramide (Gb 3 : Galα1-4Galβ1-4Glcβ-Cer)
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24 11 10 24 12 10 30 1 0.45m1.00m 1.00m 1.00m 0.33m 0.33m 0.33m 0.45m 1.00m 2 23% 29% 71% 67% 6% 4% n=1525 n=1137 6% +6% -4% -2% 21% 30% 5% 35% 6% 6% 11% 40% 37% 36 172 166 371 213 226 177 54 382 704 216
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CL-7IR-X (.x.5x.mm) Lead-free RS-4 (.65x7.5x.75mm) Lead-free PR- (.5x.375x.6 mm) Lead-free P.7 P.4 P.35 CL-9IRS-X (x.8x.8mm) Lead-free RS-47 (3.x4.5x.mm) Lead-free CPT-3 (3.xx.mm) Lead-free P.8 P.6 P.37
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2008/02/05 Isoiso FACSCalbur 488nm FL1 530/30 FITC GFP alexa488 FL2 585/42 PE Cy3 PI FL3 650LP Percp PC5 PI PE-C 7 Red APC Falcon BD Calibrite 3 BD No.340486 DNA QC particles BDNo.349523 Flow-Cheak(10um)
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1 1 36 223 42 14 92 4 3 2 1 4 3 4 3429 13536 5 6 7 8 9 2.4m/ (M) (M) (M) (M) (M) 6.67.3 6.57.2 6.97.6 7.27.8 8.4 5 6 5 6 5 5 74 1,239 0 30 21 ( ) 1,639 3,898 0 1,084 887 2 5 0 2 2 4 22 1 3 1 ( :) 426 1500
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