Fluorescence Imaging for Life Sciences forum: topic
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Which imaging techniques do you use?
Noah Gray
Monday, 18 February 2008 17:43 UTC
Well, I was disappointed that there hasn’t been a forum topic posted to this group yet, so I figured that we could start with one that everyone can answer.
During mu post-doc, I was conducting in vivo imaging of tagged proteins in the superficial neurons of mouse somatosensory cortex. These neurons usually expressed mRFP to fill the cell and reveal morphology, as well as a GFP-tagged (or photoactivatable GFP-tagged) synaptic protein, like PSD-95 or Homer1b. Although I sometimes used adults, I mainly imaged with younger animals, P8-P30.
In addition, I conducted bulk-loaded Ca2+ imaging, again in somatosensory cortex. We were looking at the responses of cells, based on somatic Ca2+ transients, after deflecting a single whisker.
The animals were all anesthetized in my work, but colleagues of mine were beginning to master these techniques in awake animals, with behavior not too far off.
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Sometimes it just takes one person to get things started. :)
I’ve used live laser-scanning confocal to perform FRAP studies on drosophila larval neuromuscular junctions expressing GFP-tagged NSF2 and actin.
I’ve also used a spinning disk confocal to image the response of various GFP-tagged markers to hypertonic shock and to anti-diuretic hormone
Lately I’ve been playing around with our TIRF microscope to image exocytosing vesicles… this machine is really temperamental!
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