Multi-functional optical imaging system (다기능성 광학 이미징 시스템)

STORM

이미징 모드 (Imaging mode)

  • 광시야 형광 현미경, Conventional Fluorescence Microscopy
  • 전반사 형광 현미경, Total Internal Reflection (TIRF) Fluorescence Microscopy
  • 단분자 현미경, Single Molecule Localization Microscopy (SMLM)
    – STORM: 25/50 nm (횡방향/종방향 분해능) for organic dyes
    – PALM: 40/100 nm (횡방향/종방향 분해능) for fluorescent proteins
    – PAINT (Point Accumulation for Imaging in Nanoscale Topography)

사양 (Specification)

  • Nikon’s Ti2-E platform with laser lines of 405/488/560/642 nm
  • Objectives: 100x/1.45 (oil), 100x/1.49 (oil, TIRF), 60x/1.27 water, 20x/0.75
  • Camera: EMCCD (Andor’s iXon Ultra 888; 1024×1024 pixels)
  • Image acquisition: NIS-Elements AR and Home-built software

장비 사용 문의

  • 장비 위치: 서울대 융합과학기술대학원(광교) D동 B106호
  • 담당자: 김정민 (jeomik@snu.ac.kr, (+82) 31-888-9082)
  • 장비 사용료 안내

Laser sources

  • Laser diode system: 405 nm CW, 50 mW (OBIS 405 LX 50 mW, Coherent)
  • DPSS laser 405 nm CW, 250 mW (LSR405NL-250)
  • DPSS laser 405 nm CW, 500 mW (LSR405CPD-500)
  • DPSS laser: 488 nm CW, 200 mW (Sapphire 488-200 OEM, Coherent)
  • DPSS laser: 488 nm CW, 200 mW (Sapphire 488 HP, Coherent)
  • Fiber laser: 560 nm CW, 1.0 W (2RU-VFL-P-1000-560-B1R, MPB Communications)
  • Fiber laser: 642 nm CW, 1.0 W (2RU-VFL-P-1000-642-B1R, MPB Communications)
  • Fiber laser: 642 nm CW, 300 mW (VFL-P-300-642-OEM1-B1R, MPB Communications)
  • Ti:Sapphire femtosecond laser: 680-1080 nm (tunable), 140 fs pulses at 80 MHz, GDD compensation (Chameleon Vision II, Coherent)
  • Superluminescent diode: 880 nm, SM fiber output (SLD880S-A25+CLD1015, Thorlabs)

  • Ti:Sapphire fs laser (Chameleon Vision II)

High-end cameras

  • sCMOS camera (Andor’s Sona 4.2B-6, SONA-4BV6U): 2048×2048 pixels, 6.5 µm pixel size, >95% peak QE, 43 fps (12/16 bit; USB 3.0)
  • sCMOS camera (Photometrics’ Prime BSI express): 2048×2048 pixels, 6.5 µm pixel size, >95% peak QE, 43 fps @16 bit (or 95 fps @11 bit; USB 3.2)
  • sCMOS camera (PCO’s Panda 4.2): 2048×2048 pixels, 6.5 µm pixel size, >80% peak QE, 40 fps (16 bit; USB 3.1)
  • EMCCD camera (Andor’s iXon Ultra 888, DU-888U3-CS0-BV): 1024×1024 pixels, 13 µm pixel size, >95% peak QE, 26 fps (16 bit; USB 3.0)
  • CMOS camera (Basler’s a2A4504-18umBAS): 4504×4504 pixels, 2.74 µm pixel size, 66% peak QE, 18 fps @12 bit)

Optical modulators

  • AOTF (97-03151-01, G&H): CA 2.5 mm, 400-650 nm, 55-110 MHz
  • SLM (PLUTO-2-VIS-096, Holoeye Photonics AG): Reflective LCOS (phase only), 1920×1080 pixels, 8 µm pixel pitch, ~2π phase range (@650 nm; 8bit), 95~98% reflectivity (dielectric mirror coating, 450-650 nm)
  • DMD (E4750RGBLC, EKB Technologies): 1920×1080 pixels (0.47″), 5.4 µm mirror pitch, 1.44 kHz rate (binary), visible (420-700 nm)
  • Deformable mirror (DMH40/M-P01, Thorlabs): 40 actuator array, Ø14.0 mm pupil diameter, protected sliver coating (0.45-20 µm), 4 kHz rate (USB2.0)

Optical scanner

  • Galvanometer scanner: non-resonant (to be updated)

Precision Actuator (Piezo)

  • PIFOC Objective scanning system (PD72Z2CA0, Physik Instrumente): 250 µm travel, 1.5 nm resolution (closed loop)
  • PIFOC Objective scanning systems (Physik Instrumente): 400 µm travel, closed loop control

Microscope objective lens

High-end objectives!
  • 100x/1.45, oil immersion (Nikon CFI Plan Apo Lambda 100x oil)
  • 100x/1.45, oil immersion (Nikon CFI Plan Apo Lambda D 100x oil)
  • 100x/1.49 TIRF, oil immersion (Nikon CFI SR HP Apo TIRF 100xc oil)
  • 100x/1.35, silicone oil immersion (Nikon CFI SR HP Plan Apo Lambda S 100xc sil)
  • 60x/1.27, water immersion (Nikon CFI SR Plan Apo IR 60xc WI)
  • 100x/1.4 (inf/0.17), Olympus UPLSAPO100XO
  • 50x/0.95 (inf/0), Olympus MPlanApo50x
  • 20x/0.75 (inf/0.17), Nikon CFI Plan Apo Lambda
  • 20x/0.40 (inf/0), Olympus LMPlanFl
  • 100x/1.5 TIRF, oil immersion (Olympus UPLAPO100XOHR)
  • Calico AMS-AGY v1.1 (inf/0, f = 5 mm, NA 1.0)

Instrument

  • Hand-held refractometer (PAL-RI, Atago): refractive index from 1.3306 to 1.5284
  • Optical Microscopy Educational Kit (EDU-OMC1/M, Thorlabs)
  • Compact spectrometer (CCS200/M, Thorlabs): 200-1000 nm (wavelength range), <2 nm FWHM (@633 nm, spectral accuracy)
  • Digital oscilloscope (MSO5104, Rigol): 100 MHz BW, 4 CHs (Analog), 8 GSa/s, 2 CHs (Func. Gen.)
  • CO2 incubator (Heraeus BB15, Thermo Scientific): 150 L