FIJI (ImageJ): Background Correction of Fluorescence Images

[@Patrick-bv7jp]

very helpful tutorial!

[@yanzheliu9092]

Many thanks!!!!!💯

[@asmaelaouina7047]

Very informative video. Many thanks for your efforts!

[@johanna.m.dela-cruz]

My pleasure. Thanks for watching. 😊

[@mathewduguay2898]

Hello! Can you do any kind of quantitative analysis after applying these methods, as long as the methods are applied uniformly over all samples?

[@johanna.m.dela-cruz]

Hi Matthew. When quantification is required, background correction is good.

[@johanna.m.dela-cruz]

Remember, you can always do better!

[@tomaszuspienski4833]

Hi very comprehensive video, short and clear. Is an image processed in this way suitable for fluorescent measurement and for comparison with another image processed in the same way?

[@johanna.m.dela-cruz]

Hi @Tomasz. If images are acquired the same way (same acquisition parameters), post-processing prior to measurements are best done in a similar way. Thanks for taking the time to give me your feedback.

[@felinetropical8822]

Can I perform deconvolution to an image if I had processed it with "background subtraction" or vice versa?

[@johanna.m.dela-cruz]

Hi Feline. Deconvolution works best if images have high signal to noise ratio. Background subtraction does help to achieve this.

[@mathewduguay2898]

Great video! Are "background correction" and "background subtraction" synonymous? I always thought we needed those special coloured slides to image and work from.

[@johanna.m.dela-cruz]

Hi @Mathew Duguay. Thanks for watching. Background correction and subtraction are used synonymously. They both mean correcting image defects mainly caused by uneven illumination in fluorescence microscopy post-acquisition. Fluorescent plastic slides are generally used during the acquisition process as a quick check on the uniformity of illumination and whether it is centered.

[@mathewduguay2898]

@@johanna.m.dela-cruz Oh I see. So the coloured slides are not imaged or used in processing. That's part of what I was confused about, I have these coloured slides but no procedure for correction or subtraction mentioned them. Thanks for clearing that up :)

[@huanyili9019]

Hey, I like your video very much. How can I find some datasets to practice ?

[@johanna.m.dela-cruz]

Hi HuanYi Li. Fiji/ImageJ has sample images (File>Open Samples) that you can use for practice. You can also check out other resources at www.cellimagelibrary.org/home.

[@cristhiancampob]

Hii Omg please can u help me🥹 i cand do it. Even i cant download the ImageJ in the Mac. This week i have a presentation, please can u help me? Regards from Mexico

[@johanna.m.dela-cruz]

Hi Cristhian. Send me an email. I’ll help you. By the way, you can download Fiji from here - fiji.sc/#. There’s a version for Mac.

[@cristhiancampob]

@@johanna.m.dela-cruz Wow. In first place Thank u very much for answer me. Wow. 2. Please give me ur mail account

[@johanna.m.dela-cruz]

@@cristhiancampob Check my “About” info in my channel.