Zeiss Axio Observer X.1 SD & TIRF

 

The zeiss Spinning Disk/TIRF system is a custom built instrument.  Built upon an Axio Observer X.1 body it is equipped with a Yokogawa Spinning disk CSU-X1 module providing confocal imaging at rapid frame rates captured on an EM CCD camera (Quant EM) ideally suited for low light imaging. Attached to a different microscope port is a highly sensitive high frame rate sCMOS camera (Orca Flash 4) which can be used for standard epifluorescence, rapid ratiometric imaging utilising a fast excitation switching light source (Lambda DG4) or acquire fluorescence images using TIRF illumination. 
Microscope body:
Inverted
Motorised Stage:
Yes
Temp. Control:
Full Enclosure
CO2:
Stage insert
Software:
Zen 2012

 


Common Applications


Epi-Fluorescence Conventional widefield fluorescent imaging.......
TIRF  Total Internal Reflection Florescence microscopy is a technique by which optical sectioning is achieved by restrictuing fluorphore excitation to within a few nundred nanometer of the glass/water interface. This technique is very useful for observing the basel membrane of adherent and migratory cells, as well as observing single molecules on glass surfaces.
Spinning Disk Spinning Disk microscopy allows fast confocal-like optical sectioning. By utilising multiple pinholes in a rotating disk several points can be scanned in parallel enabling rapid aquisiations. Especially useful when trying to observe fast biological processes, such as vesicle transport

Ratiometric Calcium Imaging

Calcium imaging is a technique by which a flourophore whoes properties are sensitive to calcium concentrations is used to report on dynamic changes in intracellular calcium levels. Simple intensity based techniques can suffer from several complications such as florescence bleaching and changes in cell morphology. Ratiometric approaches solve this issue but require very rapid instrumentation to yield accurate results. This instrument is equipped with a rappid light source switcher (lambda DG4) and a scientific scCMOS camera whose read out time is under 1ms.


 


Objective Lenses


MAG N.A CORRECTIONS IMMERSION W.D MISC.
20x 0.4 LD Plan Neofluar AIR 7.9mm  
40x 0.75 EC Plan Neofluar AIR 0.71mm DIC
40x 1.3 EC Plan Neofluar OIL 0.2mm DIC
63x 1.3 LCI Plan Neofluar Water/Glycerol 0.17mm DIC
63x 1.4 Plan Apochromat OIL 0.19mm DIC
100x 1.46 alpha Plan Apochrmat OIL 0.11mm TIRF, DIC, (UV)
* Click on objective magnification for manufactures specification sheet. 

 


Light Sources


Lasers

Diode: 405nm

Argon ion: 458nm 488nm 514nm

DPSS:561nm

Xenon  Lambda DG-4: 300 Watt Xenon arc bulb

 


Filters

 

Epifluorescence & TIRF
Name Excitation Dicrhoic Emission
21 HE FURA

BP 325 - 355

BP 379 - 395

BS 410 BP 465 - 555
43 HE DsRED BP 538 - 562 BS 570 BP 570 - 640
74 HE GFP/mRFP

BP 464 - 496

BP 552 - 577

BS 505

BS583

BP 510 - 540

BP 588 - 645

 

Spinning Disk
Dichroics Emission Filters

405/488/532/647

457/514/647

BP 450/50

BP 485/30

BP 525/50

BP 629/62


 


Detectors


Detector Pixel Format SeNsor Description
Orca Flash 4 V2 2048 x 2048 sCMOS ???????
QuantEM 512SC 512 x 512 emCCD ?????????

 

Test Document

 

Contact People

Dr Renee Whan

Dr Renee Whan

Position: Senior Lecturer and Head
Facility: Biomedical Imaging Facility

Full profile

Dr Michael Carnell

Dr Michael Carnell

Position: Research Associate
Facility: Biomedical Imaging Facility

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Dr Alex Macmillan

Dr Alex Macmillan

Position: Research Scientist
Facility: Biomedical Imaging Facility

Full profile