live cell imaging
Great news we are currently looking for a new honours student for 2016.
The title of the project is “Developing novel biosensors to monitor DNA damage in cancer cells”.
Its a very exciting new project incorporating cutting edge microscopy and fluorescent biosensors.
If you think you have what it takes and are interested please feel free contact myself, or UNSW SoMS.
For more information on the UNSW honours program please visit: http://medicalsciences.med.unsw.edu.au/students/soms-honours/
Below is an example of the images that will be created during the project.
I often get asked how to uses Thresholds to measure things in Image J.
Below are some of the Basic Steps for using Thresholds:
- Open your image and duplicate it (Image>Duplicate)
- On the duplicate go to Image>Adjust>Threshold
- Play with the sliders until all of your cells are red.
- Click ‘Apply’
- You should now have a ‘binary’ black and white image
- Now go to menu Process>Binary and select ‘fill holes’
- You may also want to select erode, dilate, open or close to optimise the binary image so that you have nice solid filling of your cells.
- Now go to menu Analyse>Set Measurements. Select all the things you want to measure.
- Critical steps: make sure that you select your original image (not the binary) in the ‘Redirect to:’ pull down Menu
- Also make sure the ‘Limit to threshold’ checkbox is ticked and also tick the ‘Add to overlay’ and ‘Display label’.
- Click ok to close the ‘Set Measurements’ box.
- Now go to Analyse>Analyse Particles
- Here you will need to play around with the size and circularity settings (bit of trial and error) in order to get accurate identification of your cells or ROIs. I suggest making duplicates before you start so that you can quickly try different things to see which one works best.
- Make sure you have the Display results tick box selected.
- Once you click ok you should have a the measurements box appear with all your measurements for each cell.
- You can copy and paste these into Excel or what ever program you like to use.
- Go get a coffee and cake you deserve it!
Video Posted on
Here is a slide that I made and often use at the beginning of my talks to briefly explain what mitosis is. The images are taken from a live cell that we have labelled with fluorescent probes to highlight the DNA (red) and microtubules (green).
Great news for those who cannot afford the very price commercial alternatives for 3D analysis of microscope images.
You can download the software here
And you can read the original research article published in Nature Protocols here
Video Posted on Updated on
We have been busy constructing some new cell lines that stably express multiple fluorescently tagged proteins so that we can visualise different aspects of mitosis in real-time. The results have been stunningly beautiful.
You can check them out on our Vimeo Channel