SD2 - Andor Spinning Disc FRAPPA

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Suitable for confocal imaging, photobleaching and -activation of DAPI,EGFP, mCherry, Cy5 with high temporal and spatial resolution.
2012 09 10 SD2 FRAPPA.jpg


Logo MPI-CBG.jpg
MPI-CBG-Lmf-logo-big.pngMPI-CBG LMF (MPI-CBG, 038B)



microscope Olympus - IX81, Inverted stand, motorized with ZDC (hardware autofocus), 2x Optovar, Prior NanoScanZ, Prior ProScanIII xy scanning stage
Olympus UPlanSApochromat 10x 0.4
Olympus UPlanSApochromat 60x 1.2 W
Olympus UPlanSApochromat 60x 1.35 Oil
Olympus UPlanSApochromat 100x 1.4 Oil
Olympus UApochromat 40x 1.15 W
Transmitted Light (LED)
Fluorescence (HBO)
Laser Diode 405 nm
Laser DPSS 488 nm
Laser DPSS 561 nm
Laser DPSS 640 nm
detection [[detection::Spinning disc scan head Yokogawa CSU-X1 (5000rpm):
dichromatic mirrors:
Quad band T-405/488/568/647
Triple band T-405/488/561
Single band T-488
physical pinhole radius: 25um, pinhole spacing: 250um; back-projected pinhole size: 0.4um, back-projected pinhole spacing: 4um (with 60x objective, 1x optovar)
Andor Dual camera port with two cameras:
Andor iXon EM+ DU-897 BV back illuminated EMCCD; dexel size of EMCCD chip: 16um
Andor Clara cooled interline CCD; dexel size of chip: 6.45um
note there are two additional lenses in the optical path: 1.0x before the CSU and another one after the CSU: a 1.97x in front of iXon or a 0.95x in front of Clara
image pixel size for use with iXon (measured Sept. 8th 2012): with 100xoil objective: 0.081um/px (optovar 1x), 0.041um/px (opt. 2x); with 60xW obj.: 0.136um/px (opt.1x), 0.068um/px (opt 2x); 10x: 0.811um/px (opt.1x), 0.407um/px (opt.2x)
pixel size with Clara (measured Sept. 8th 2012): 100x oil objective: 0.068um/px (opt 1x), 0.034um/px (opt2x); 60xW obj.: 0.114um/px (opt.1x), 0.057um/px (opt. 2x); 10x obj: 0.68um/px (opt. 1x), 0.34um/px (opt. 2x)

old values:
projecting lens in front of iXon: was 2.475 before installation of TuCAM on Sept 4th 2012; was 2.39x before removal of DualView on Dec 21st 2011; was 2.067x before addition of the dual filter wheel on Apr 28th 2011, was 1.9x before addition of dual camera switch on June 30th 2010
projecting lens in front of Clara: was 1.285x before installation of TuCAM on Sept 4th 2012; was 1.28x before removal of DualView on Dec 21st 2011; was 1.2x before addition of the dual filter wheel on Apr 28th 2011

image pixel size for use with iXon (measured Jan 2012): with 100xoil objective: 0.0646um/px (optovar 1x), 0.0322um/px (opt. 2x); with 60xW obj.: 0.108um/px (opt.1x), 0.054um/px (opt 2x); 60x oil obj.: 0.1074um/px (opt. 1x), 0.0537um/px (opt. 2x); 10x: 0.648um/px (opt.1x), 0.324um/px (opt.2x)
pixel size with Clara (measured Jan 2012): 100x oil objective: 0.05um/px (opt 1x), 0.025um/px (opt2x); 60xW obj.: 0.084um/px (opt.1x), 0.042um/px (opt. 2x); 60xoil obj.: 0.0834um/px (opt.1x), 0.0417um/px (opt. 2x); 10x obj: 0.502um/px (opt. 1x), 0.251um/px (opt. 2x)
image pixel size used with iXon (measured Ju;y 15th 2011): with 100x: 0.067(was 0.077um before Apr 2011); with 60x objective: 0.111 (was 0.129 before Apr 2011) (was 0.14um); 60x obj. and 2.0x optovar: was 0.0645um (was 0.07um); 40x obj.: was 0.194um (was 0.21um); 40x obj. and 2.0x opt.: was 0.0967um (was 0.11um)
pixel size with Clara (measured July 15th 2011, all with optovar 1x): 100x objective: 0.05um/px, 60x obj.: 0.084um/px, 10x obj: 0.511um/px
reflectors Sutter filter wheel A:
position 0: 440/521/607/700 Quadband, 1: razor edge 488LP, 2: BL700/75, 3: razor edge 568LP, 4: Edge Basic longpass 635 LP, 5: BrightLine 445/45, 6: open, 7: polarizer for DIC, 8: 525/30, 9: BrightLine HC 607/36
Sutter filter wheel B:
position 0: empty/open, position 1: BL 470/22; positions 2 - 9: empty/open
microscope filter turret:
position 1: empty, 2: DIC, 3: dual GFP/mCherry ET, 4: EGFP ET (X 470/40; D 495LP; M 525/50), 5: Tx Red ET (X 560/40; D 585LP; M 630/75, 6: DAPI HC (X 377/50; D 409LP; M 447/60)
features objectives 40x 1.25 SIL and 60x 1.3 SIL are shared objectives and need to be booked separately via the LMF booking database!
software [[software::iQ 3.5 (with Python) iQ3 user guide

connections: COM1 - stage; COM2xx - lasers; COM3 - CSU; COM4 - Sutter FW; COM5 - Olympus IX 81; COM6 - FRAPPA; AOTF - ALC401/501/601; DD A06/16]]

incubation Stage incubator (T) + objective heater
  • [- beam path]
  • [ advanced system check] 0

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