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

Price structure

Fixed Price Structure for Booking

The fixed annual price structure for AxioObserver Inverted Widefield usage allows unlimited booking of both the AxioObserver (Skou building) and the AxioObserver (Anatomy building). Within this structure, it is also possible for registered users to use the instruments for a quick sample check by booking a 15 min time slot.

Training Session

A training session is mandatory and is not included in the annual price. As the AxioObservers hold minor differences, the training session will be given to the most relevant AxioObserver and, if needed, supplemented with a free of charge, short add-on for the other AxioObserver.

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)

FAQ

Equipment

- How should I clean the equipment after use?

If you have used the 63x Oil and/or 100x Oil objective(s), ensure that you have gently wiped off the oil droplet with a piece of designated lens paper.
In case you accidentially contaminated air objectives with oil, please inform us immediately to ease the cleaning process for us as much as possible.
Please only use designated lens paper for wiping off oil from objects and designated tissues for cleaning of samples.
Please leave the work station neat and clean for the next user.
Ensure to cover the microscope with the designated dust cover.

- Are consumables included in the price?

Experimental consumables are not included in the price for booking of microscopes via the Bioimaging Core Facility.
Designated oil for the oil objectives can be found next to the microscope. Please only use this designated oil for the objectives.
Lens paper designated for cleaning of objects as well as designated tissues for cleaning of samples are located next to the microscope.

Data Storage

- How can I get access to my data?

After using a microscope or a workstation, please transfer your data immediately to your personal AU drive (O:) and clean up your local directories on instrument PCs if you are not reusing data in subsequent experiments.
The Bioimaging Core Facility staff will provide support to non-AU users for exporting data.
Use of USB memory sticks or external hard drives is for security reasons not allowed.

- Does the Bioimaging Core Facility regularly back up the user data?

The Bioimaging Core Faility is not responsible for your data. DATA ON LOCAL PC DRIVES WILL BE DELETED ON A REGULAR BASIS in order to keep the systems performing optimally.

Please ensure that you transfer your data to your personal AU drive (O:) after each session.

- May I use my personal USB memory stick or external hard drive for data storage?

No, any use of USB memory sticks or external hard drives is for security reasons not allowed. Usage of USB memory sticks may lead to suspension of the user license.