FD-FLIM

 

FLIM Location: LCAM, Room A2.34
Contact person: staff-Dorus
2nd Contact person: staff-Joachim
Techniques: (Semi-)Widefield imaging
Confocal imaging (using spinning disk)
Spectral imaging microscopy (SPIM)
Fluorescence Lifetime Imaging Microscopy (FLIM)

Total Internal Reflection Microscopy (TIRF)
Booking: The FLIM microscope can be booked via the webcalender and is only available for trained users. For first time-users contact Dorus Gadella (+6259).
Lasers: 405 nm (diode laser)
442 nm (diode laser) 

488 nm 
514 nm
532 nm
568 nm 
For more info about laser safety, contact Ronald Breedijk (+7860)
Description: The Zeiss Axiovert widefield microscope is equiped with insturments for frequency-domain FLIM or spectral imaging. The setup can be extended with a spinning disk device for confocal measurements. Mercury lamp filters: 365 nm
405 nm
436 nm
470 nm
500 nm
546 nm
577 nm
Objectives: 10X (Plan NeoFluar NA 0.3 air)
20X (Plan NeoFluar NA 0.5 air)
40X (Plan NeoFluar NA 1.3 oil)
63X (Plan Apochromat NA 1.4 oil)
63X (AchroPlan NA 0.95 water, no coverslip, long WD)
100X (AlphaPLAN NA 1.45 oil) 
Microscope: Zeiss Axiovert 200M
Detection filters: download filter-scheme (not available yet)
Beamsplitters:
CFP
GFP
YFP
RFP
80/20% mirror
Calibration Cube
Emission filters:
CFP bandpass
GFP bandpass
YFP bandpass
RFP bandpass
400 nm longpass
430 nm longpass
460 nm longpass
500 nm longpass
525 nm longpass
570 nm longpass
600 nm longpass
Accessoires: Perkin Elmer UltraVIEW Spinning disk unit
Image intensifier (Lambert Instruments)
Cooled CCD camera (Coolsnap HQ)
Spectrograph (400-700nm) Imspector
Cooled CCD camera (Hamamatsu ORCA II) 
Software: LCAM Image J macro to create colored-FLIM images
Manuals: LCAM FLIM Start-up manual (not avialable yet) 
LCAM FLIM Laser manual
LCAM user rules 

LCAM startup procedure 405 laser
LCAM multifrequency-FLIM procedure

LCAM EB calibrationprocedure/image processing

 

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