Hi all,
Recently, I am trying to measure LTP signal, but I cannot detect the response.
I sliced the brain horizontally with vibratome. For slice, i used ice-chilled aCSF (0.5 mM, CaCl2), and the slices were recovered in normal aCSF (2 mM CaCl2) for 1-2 hrs.
After recovery, I transferred the slice to recording chamber where is perfused warm aCSF (> 2ml/min of speed).
For stimulation I used concentric bipolar electrode (FHC, pencil form), and the stimulation was applied for 0.1 ms with various strength. for recording, I used HEKA EPC 10 and patchMaster, and the electrode was assembled with glass pipette filled (1-3 MOhm) with aCSF. current clamp mode was used to measure the change of voltage with holding current as 0.
however, still I could not get any signal. please give me some advice for this.
Thanks!
Hello, Max!
I was trying to record in CA1 region by stimulating SC fibers
The age of mice were about 4 weeks old.
I added the pictures!
Best,
HanGyu
Han-Gyu Bae
Hi,
were you able to figure out the problem?
if so, I"ll be thankful if you may share what was the problem and how did you solve it, since I'm facing the same issue.
thank you
Hey, Samaa Zidan,
I finally figured out the reason why i cannot detect the signal.
It was quality of slice. when I was doing the slicing, i used vibratome with blade speed of 0.26 mm/s and 1 of amplitude. but some how when i increased the blade speed to 1.2 mm/s, the signal showed up. i think when I used slow speed the SC fibres were exposed to vibration too long so most of them physically damaged. but when i increased the speed, maybe, the exposure time to blade vibration got shorten and less fibres were damaged. I think this is the why many people prefer to use chopper instead of vibratome for LTP experiments. Try to increase you blade speed. let me know if you also can succeed in detecting the signal.
Regards,
HanGyu
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