BQWANG at MACC.WISC.EDU
Thu Aug 6 17:48:00 EST 1992
I wrote a post to Neuroscience which said:
>I am trying to use Lucifer Yellow (LY) filled glasselectrode to do
>intracellular recording, and then inject LY into the cell to label
>it. I find that it's difficult to keep the cell for a long time.
>Would anyone please share your experience with me?
I am grateful that I got many suggestions.
>>>Frank: 1> make the electrode respect the size of the cell; 2>
table being isolated from vibration. 3> high intensity UV will kill
the cell. 4> use a electrode of high resistance (10-15 MegOhms) and
a sharp shank.
>>>Ken Scholz: 1> It depends on the type of cells. 2> Lucifer
Yellow is toxic to invertebrate neurons. (vertebrate cell?)
>>>Andrew Bangham: (used to work on B-cells successfully) the
crucial part is the very long, thin, high resistance (3-7 10^8
>>>David Parsons & Scotty: asked for more information about animal,
cell type, electrode tip, method of injection.
Here are some more information of my experiment. I use hemisected
spinal cord of frog, I don't know for sure the size of the dorsal
horn neuron. A scholar in my lab used 100-150 MegOhms glass
electrode ( filled with 3M KCl), and did successful recording. But
LY (8% dissolved with 0.5M LiCl) makes the resistance too high to
do recording. If I make a electrode with low resistance (filled
with LY), the tip of electrode will too big to fit the cell. I am
still trying to find a right electrode whose resistance wouldn't be
too high and tip wouldn't big. I think it's tough but possible.
And, I use electric pulses to inject LY after recording.
Any more advice is appreciated.
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