First flight tests, Trehalose again…
on Thursday, November 1st, 2012 7:33 | by Christine Damrau
Chrisi’s day…
1 Collected virgins for our ill post-doc, made some crosses:
d42-GAL4 males x tubGAL80-UAS-PKCi and x CS
d42-chaGAL80 males x tubGAL80-UAS-PKCi and x CS
2 Buridan: Prepared colored food (10mg/ml, 2.5ml total in the very little vials, ~ 5 flies in each vial). Denise tested in the afternoon females in the Buridan. Data (4n now) will be shown in the lab meeting.
3 Imaging Made a new cross for the Imaging-project:
TDC2-GAL4 males x UAS-GCaMP3(2)
replaced Neloy’s flies by flies from stock
Florian’s larval preparation (good!) showed that GCaMP-expression is much less than GFP-expression. They hope for the homozygous line.
4 Flight: Performed flight experiments with the tbh,TDC,UAStbh flies. Raw data, see below. Thinking about analysis (probably tomorrow)
Protocol: glued the day before, sugar in wheel chambers, little EVIAN-water.
5s filter on the legs before starting, aspirated, stopped when no response 3 times in a row, stopped time when 30min, stopped after 15 aspiration trials
| date | fly | genotype | number_aspiration | time_to_first_stop | duration_of_flights | remarks |
| 01.11.2012 | 1 | 12 | 4.56 | 4.56,0.22,0.30,0.2 | ||
| 01.11.2012 | 2 | 2 | 49.15 | 49.15,14.42 | ||
| 01.11.2012 | 3 | 16 | 0.03 | 0.03,0.15,3.06,1.56,0.3,0.14,4.58,0.22 | ||
| 01.11.2012 | 4 | 3 | 0 | 0 | ||
| 01.11.2012 | 5 | 40 | 0.05 | 0.05,0.01,0.02,0.01,0.01,0.02,0.02,0.01,0.02,0.03,0.02,0.02,0.01,0.01,0.01,0.01,0.01,0.04,0.09,0.01,0.02,0.01,0.11,0.01,0.01,0.01,0.01,0.01,0.02,0.01,0.03 | ||
| 01.11.2012 | 6 | 3 | 0 | |||
| 01.11.2012 | 7 | 13 | 3.01 | 0.01,0.01,0.01,0.01,0.01,0.02,0.01 | ||
| 01.11.2012 | 8 | 15 | 0.01 | 0.01,0.04,0.03,0.02,1.56,0.15,0.02,1.46,3.26,1.15,0.46,6.45,2.56,0.06,12.59 | ||
| 01.11.2012 | 9 | 15 | 0.01 | 0.01,0.01,0.28,0.1,1.5,1.29,0.40,0.29,1.34,0.54,0.22,0.37,0.31,0.54 | ||
| 01.11.2012 | 10 | 9 | 0.01 | 0.01,0.01,0.01,0.01,0.01 | ||
| 01.11.2012 | 11 | 12 | 0.01 | 0.01,0.01,0.01,0.01,0.01,0.01 | ||
| 01.11.2012 | 12 | 3 | 0 | |||
| 01.11.2012 | 13 | 4 | 0.06 | 0.06,0.5,0.06,38.2 | ||
| 01.11.2012 | 14 | 3 | 0.19 | 0.19,0.33,0.31,30 | ||
| 01.11.2012 | 15 | 15 | 3.38 | 3.38,2.43,0.47,0.36,4.06,0.39,1.25,0.35,0.52,1.07,0.22,0.23,0.27 | ||
| 01.11.2012 | 16 | 3 | 0 | |||
| 01.11.2012 | 17 | 3 | 7.16 | 7.16,23.27,30 | ||
| 01.11.2012 | 18 | 3 | 0 | |||
| 01.11.2012 | 19 | 6 | 1.33 | 1.33,14.57,9.17,30 | ||
| 01.11.2012 | 20 | 1 | 18.20 | stopped the expriment | ||
| 01.11.2012 | 21 | 3 | 2.50 | 2.50,2.18 | stopped the expriment |
5 Trehalose: Found time to try out Gérard’s idea (no pigment in the probes to not disturb the NanoDrop measurement). 4 groups: 10 flies with or without heads, 50 flies with or without heads. Hilde showed me how to remove heads with liquid nitrogen and vortex. The other flies (with heads) were frozen at -20° as usual. Trehalase incubation over night. Tomorrow glucose oxidase and measurement
Category: Biogenic Amines, buridan, flight, trehalose measurement
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