Fluorescence detection in living cells and ways to improve your image

Sponsored by: Life Technologies
Session ends: December 13th, 2013, 3:00pm MST
Answers by: Timothy Fawcett, Ph.D., Director, BioTechnical Institute of Maryland (BTI)


Fluorescence imaging of living cells can provide important data regarding the function and localization of proteins and other bio-molecules within a cell or tissue. These images give insight into fundamentally important biological processes and improve our knowledge about transient interactions we might not be able to detect otherwise. An added benefit is some simply remarkable pictures of colorized cells which are just fun to look at. Although in theory, fluorescence microscopy is simple, obtaining suitable images is difficult. Problems with cell health can occur due to long incubations in D-PBS in an attempt to reduce auto-fluorescence. Cell death due to light intensity or photo-bleaching can be problematic and need to be overcome. If you are having problems with signal:noise or cell health or obtaining the best image possible now is your chance to ask the expert.

Ask the Expert Image

This session is sponsored by
Life Technologies

Don’t miss this chance to have your cell imaging questions answered and also get the chance to win a free set of six spray bottles for your lab, courtesy of Life Technologies!


Questions & Answers

What methods are best for looking at intracellular events?

Hi I have to tell you there are many methods and probes for looking into intracellular events. I could not do the topic justice in a short answer. Here is a link to the Molecular Probes Handbook which will have all the answers to your questions. http://www.lifetechnologies.com/us/en/home/references/molecular-probes-the-handbook.html Have a look.» Read More

What do you recommend for imaging of non-adherent cells?

You can do imaging on suspension cells. If you are using phase-contrast you won’t get much since the light reflects off of the cells in suspension, but you can get images using other types of microscopy. Confocal and fluorescent imaging is possible. Often fixations are necessary. The trick is using an ultra-thin layer of cells […]» Read More

What kind of system would you recommend for live-cell imaging? Would you use live-cell incubation chambers or something else. Also what are the advantages of live-cell imaging?

There are lots of good chamber systems for live imaging. If imaging is occurring for a few hours you might be able to get away without an incubation chamber but if your incubations are longer a more sophisticated system would be necessary. Here is a good website from Nikon that talks about them (http://www.microscopyu.com/articles/livecellimaging/culturechambers.html). Obviously […]» Read More