culture: coolio25020 age: 13 DIV temperature: 30C ROI: 01 white LED level: 15 (trying to use super low light so as not to activate ChR2 in any way) blue LED level: 99 exposure: 50ms x-position: 0.89837 y-position: 0.34989 z-position: 37um positioned over center electrodes recording for 5min, with 10s before and after weirdness on 37 (maybe 17 also) imaging notes: coolio25025_ROI-01_quick-test_exp-50ms_LED-15_1 --------------------- num_frames: 350 total time: 30.4586 s corresponding MCD file: none no burst in this recording. trying again. imaging notes: coolio25025_ROI-01_quick-test_exp-50ms_LED-15_2 --------------------- num_frames: 350 total time: 30.4722 s corresponding MCD file: none no burst in this recording. trying again. imaging notes: coolio25025_ROI-01_no-drug_ChR2-stim_2 --------------------- num_frames: 4100 (aborted early--didn't see frame, but we can get that from filenames) total time: 309.158s corresponding MCD file: coolio25024_ROI-01_no-drug_ChR2-stim_0006.mcd stimulation definitely evoked a bit burst (on MEA) here at 1min. there was an error (circular buffer) at the end (309.158s) just before the camera light turned off. I'm gonna just go ahead b/c it was just a second away from finishing and that's okay. imaging notes: coolio25025_ROI-01_no-drug_no-stim_1 --------------------- num_frames: 4100 total time: 339.5328 s corresponding MCD file: coolio25020_ROI-01_no-drug_no-stim_0002.mcd ADDED CNQX AT 12:25AM doing 10min 30 sec recordign as usual. channels 17 and 27 as usual are the first to show crap Good! no bursts. I used CNQX aliquots from the 3ul and 6ul (instead of 12ul) batch... wonder if that made a different imaging notes: coolio25025_ROI-01_CNQX_no-stim_1 --------------------- num_frames: 4100 total time: 340.072 s corresponding MCD file: coolio25020_ROI-01_CNQX_no-stim_0002.mcd imaging notes: coolio25025_ROI-01_CNQX_ChR2-stim_1 --------------------- num_frames: 4100 total time: 339.5984 s corresponding MCD file: coolio25020_ROI-01_CNQX_ChR2_0002.mcd YES! stimulation evoked a huge burst. it was hard in the others b/c the culture was bursting so much (even in CNQX), but when activity is legitimately down ti looks like stimulation works. maybe i used a bad stock in the others... doh! failed at 200s in MCRack still should look at first part of recording though try again... imaging notes: coolio25025_ROI-01_CNQX_ChR2-stim_2 --------------------- num_frames: aborted at 2858 total time: aborted at 237.3844s corresponding MCD file: coolio25020_ROI-01_CNQX_ChR2-stim_0005.mcd YES! stimulation evoked burst again at 1min failed again when i tried to maximize the screen (I won't do that next time) imaging notes: coolio25025_ROI-01_CNQX_ChR2-stim_3 --------------------- num_frames: 4100 total time: 339.539 corresponding MCD file: coolio25020_ROI-01_CNQX_ChR2-stim_0007.mcd gah! again it stopped at 229 sec. well i have several that show a burst with blue light stim (probably). i'll make a lite version of the rack. imaging notes: coolio25025_ROI-01_CNQX_ChR2-stim_4 --------------------- num_frames: failed at 3012 frames total time: failed at 250.2755 corresponding MCD file: coolio25020_ROI-01_CNQX_ChR2-stim_0010.mcd bah, this time Micromanager failed, but MC Rack was okay. imaging notes: coolio25025_ROI-01_CNQX_ChR2-stim_5 --------------------- num_frames: 4100 total time: 339.526 corresponding MCD file: coolio25020_ROI-01_CNQX_ChR2-stim_0013.mcd yay! finally one where neither failed. still look at previous recordings since first stim was always fine (and maybe less bleaching in earlier recordings. I'll do a another CNQX-only recording... imaging notes: coolio25025_ROI-01_CNQX_no-stim_2 --------------------- num_frames: failed at 2498 frames total time: failed at 207.4921s corresponding MCD: coolio25020_ROI-01_CNQX_no-stim_0005.mcd MM and MCRack both failed! It's probably time to call it a night. I'm being greedy with wanting more data. The signal has faded a considerable amount now.