AxioObserver (Skou), Inverted Widefield, Zeiss

Rat hippocampal neurons_Annemette Boe Marnow

Porcine endothelial cells (pBECs), Annemette Boe Marnow

Rat hippocampal neurons, Annemette Boe Marnow

Porcine endothelial cells (pBECs), Annemette Boe Marnow

The AxioObserver (Skou) Inverted Widefield Microscope is easy to use while maintaining sensitivity.

Applications in brief

This inverted widefield microscope is equipped with two light sources and two corresponding detectors (cameras), allowing generation of either fluorescence images or brightfield imagages.

Its motorized stage allows automated sample finding and generation of tile scans.

In doubt if your staining worked? Book a 15 min time slot to check your sample.

Features

Brightfield and/or multispectral fluorescence
Stage mounting frames for slides (max. length 120 mm)

Which samples may I use?

Selected samples, but not limited to:

fixed tissue sections or fixed cells
brightfield (e.g. H&E)
immunofluorescence (due to limtations by objectives, consider corresponding microscope located in the anatomy building for this application)
live cells in thoroughly sealed containers

Training and use

Tips and tricks

Consider bleed-through/spectral overlap when investigating potential co-localization

Ensure that the chosen fluorophores do not spectrally overlap.
The current filtercubes of this microscope pose limitations to co-localization studies. Ensure that the settings (choice of filters ect.) are set correctly to avoid false co-localization signals or consider using the corresponding microscope located in the anatomy building.

Imaging of live cells

For imaging of live cells, e.g. in petri dishes, ensure to seal the container with parafilm and to use a suitable stage to avoid spillage into the microscope and to ensure a horizontal sample.

Specifications

Full equipment name	Widefield Inverted Light Microscope, Zeiss
Objectives	5x/NA0.16 Air, Ph1, EC Plan-Neofluar; FWD 18.5 mm; cover glass thickness 0.17 mm 10x/NA0.30 Air, Ph1, EC Plan-Neofluar; FWD 5.2 mm; cover glass thickness 0.17 mm 40x/NA0.75, Air, Ph2, EC Plan-Neofluar; FWD 0.71 mm; cover glass thickness 0.17 mm 63x/NA1.25, Oil, Ph3, EC Plan-Neofluar; FWD 0.10 mm; cover glass thickness 0.17 mm 100x/NA1.30, Oil, Ph3, EC Plan-Neofluar; FWD 0.20 mm; cover glass thickness 0.17 mm
Filter qubes	DAPI/GFP/Cy3/Cy5 Excitation BP 385/30; BP 469/38; BP 555/30; BP 631/33 Beamsplitter QBS 405 + 493 + 575 + 653 Emission QBP 425/30+514/30+592/25+709/100 CFP/YFP/mCherry Excitation BP 423/44; BP 511/44; BP 591/27 Beamsplitter TBS 450+538+610 Emission TBP 467/24+555/25+687/145
Light sources	Illuminator microLED (for brightfield applications) Colibri 7 type RCBV-UV (for fluorescence applications)
Detectors (cameras)	Axiocam 305 color (for brightfield applications) Axiocam 705 mono (for fluorescence applications)
Heated stage	No
Motorized stage	Yes (tile scan possible)