I cannot obtain any images; there is no live image is on the screen and the live button is grayed out

• Check if the camera is turned on (red button = camera on)
• Ensure that the slider stick on the side of the microscope is either half way or all the way out

   

My 'Stage Navigator' has disappeared

• Find 'Stage Navigator' in the tab line, right click, choose 'Tool Window Mode'

   

My pictures disappear after acquisition

• Settings have likely been changed and not set back
• Consult the quick-guide located next to the microscope, section FAQ, p.8

   

There are visible shadows in my sticked image (vignettes). How do I fix this?

• Reduce the imaging area to increase the overlap used for stitching)
• Go to 'Live'
• Under 'Resolution', choose 'Toggle Subarray' and 'Apply to Live'
• Set an ROI to reduce the size of your image

   

My section has varying thickness or is mounted unevenly. How do I get a crisp image?

• Add multiple focus points
• Go to 'Stage Navigator'
• Choose 'High Density Focus Map'; you will see 5 preselected boxes/focus points
• Press the green arrow button to go through the focus points while adjusting each one
• If needed, add more focus points by right-clicking on the overview and choosing 'Add focus point'
• When all focus points are adjusted, start experiment

Full equipment name	BX63 Upright Widefield Fluorescence Microscope, Olympus
Camera	Sensitive Andor Zyla 5.5 (5 Megapixel, 6.5 µm pixel – 60% Quantum efficiency)
Objectives	10x, Plan Flurite 20x, Plan Apochromat 40x, Plan Flurite 60x-Oil, Plan Apochromat 100x-Oil, Plan Flurite
Filter qubes	Hoechst (blue), Ex 360-370nm, Em 420-460nm GFP (green), Ex 465-495 nm, Em 515-555nm mCherry (red), Ex 540-580nm, Em 605-665nm Cy5 (far red), Ex 590-650nm, Em 663-737nm
Light source	Cooled pe300 ultra with enhanced power output 3 individual bands according to observations methods are automatically controlled to avoid bleaching

• Please leave the work station neat and clean for the next user.
• Ensure to cover the microscope with the designated dust cover.

• Consumables are not included in the price for booking of BX63 microscope via the Bioimaging Core Facility.

• 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.

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.

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.

• Obtained images may be processed and analysed using Olympus cellSens software:
  • PC2: Simple file processing and conversion of files with cellSens Entry v2.3.
  • PC1: Advanced file processing including counting and measuring as well as constrained iterative deconvolution* with cellSens Dimension v2.3

* Deconvolution is an image processing technique used to improve contrast and resolution of images. Out-of-focus light causes blur, which mathematically is represented as a convolution operation. Deconvolution seeks to reassign out-of-focus light to the correct origion. Nearly all fluorescence images can be deconvolved.

• Please also ensure to acknowledge the Bioimaging Core Facility. For guidance consult Publications & Acknowledgements.