Hi
I am try to show that an organotypic skin model I have built closely resembles the structure of human skin by performing immunofluorescence with antibodies directed against a number of antigens that should be present is specific areas of the skin.
I have performed the staining and acquired my images but now need to edit the brightness and contrast of the images and crop them to size. I have downloaded Fiji and have had a play at editing the images but am a little unsure of how far to edit them and how to be consistent between the models and the skin.
Does anyone have any advise?
Many thanks
You may be able to perform some other image analysis on the model and experimental images in order to segment and measure some features of interest, so that you can QUANTITATIVELY compare the two. I strongly recommend you look at MIPAR at http://MIPAR.us for a powerful yet intuitive image analysis package. I'd be happy to make a recipe file for your automated segmentation if you are able to provide an example image.
Tara,
the very basic editing of any stack image can be done by selecting, on ImageJ:
Image-Adjust-Color Balance
Image-Color-Channel Tool
Always have both open to play around with your images. Channels tool is crucial, under "Color" you can modify each channel (including the color), under "Composite" you can look at co-localization. Be aware that double or triple channel images can be exported (using Image-Type-RGB Color) only in Composite mode.
If you have stack images, you can either show single planes or Maximum intensity projections (Image-Stacks-Z Project).
Also, the Despekle and the Smooth features can help you cleaning up your images.
- Badges
- Science method