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wiki:documentation:monika:5-ht_experiments_pumping_freely_moving

5-HT experiments pumping freely moving

Concentrations

On 3.5cm plates add 0uM, 5uM and 10uM 5-HT to center of the plate. Use small volume (ca 50 ul). Add 20 adult worms to each plate and incubate for 10 minutes.

Image each plate with normal foraging behavior conditions for 5 minutes.

First trial: I let the 5-HT dry on the plate and then added worms. I saw no effect after 10+minutes. As a test of the serotonin, I added 10 uM direcly onto worms and immediately saw them slow down and pump. Next time: Pick worms to center of 3.5cm plate first, then add 5-HT. Image when the drop is dried.

2nd Try 18.3.2020

Pick 30 GRU101 young adult to adult onto 3.5cm plates into M9 (some bacteria are transported.) Add 20ul of M9, or 5uM or 10uM 5-HT to each plates and wait until dry. wait another 5 minutes until worms disperse and start imaging. Using 15ms exposure, Gain 10. Focus looks very good on Zeiss Axio with 32x nominal (1x actual) magnification.

(0) M9 control added at 13:40, dry at 13:44
(1) 5uM add at 13:49 - start recording at 14:03
(2) 10uM added at 13:50 - start recording 14:09

MS0001 - Control (20ul M9)
MS0002 - 5uM 5-HT
MS0003 - 10uM 5-HT

and repeat…

(0) M9 control added at 14:32, dry at 14:37, recording at 14:42
(1) 5uM add at 14:40, dry at 14:46, - start recording at 14:52
(2) 10uM added at 14:46 dry at 14:53- start recording 15:00

MS0004 - Control (20ul M9)
MS0005 - 5uM 5-HT
MS0006 - 10uM 5-HT

Analysis

All in all it looks very promising. I had 110 worms over 6 recordings. The 5-HT worms show a higher pumping rate than the control, but it is not as striking as expected. My guess is it is due to the bacteria transferred over with the worms. Repeat this with 1-2 washing steps in between and we should be good. Will also plot velocity for posterity.

wiki/documentation/monika/5-ht_experiments_pumping_freely_moving.txt · Last modified: 2020/04/21 07:02 by mscholz

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