User Tools

Site Tools


wiki:documentation:wiki:documentation:nicolina:24.4.20

Differences

This shows you the differences between two versions of the page.

Link to this comparison view

wiki:documentation:wiki:documentation:nicolina:24.4.20 [2020/04/24 17:51] (current)
nzjacic created
Line 1: Line 1:
 +====== 24.4.20 ======
 +
 +===== Objectives =====
 +
 +  * Starve INF100 at 13:00 and 16:30
 +  * Starve GRU101 at 16:30
 +  * Record as many videos of INF100 using the macroscope as possible
 +  * Lightning group meeting
 +  * Talk with Luis about Python software that would be useful for macroscope recordings
 +  * Put only 5xL4 onto maintenance plate of INF103 to already start gauging timing for egg-laying adults
 +  * Took out plate of refreshed OP50-GFP left overnight to grow at 37C
 +
 +===== Observations =====
 +
 +  * INF100
 +    * Were still at L4 when I wanted to starve them at 11:00 - waited until 13:00 to let them grow longer
 +    * Est. take about 72 hours from egg to adult ready for recording - much easier to adjust sync times this way
 +  * Macroscope testing
 +      * **ISSUES:**
 +        * Camera cannot focus on lawn and pharynx at the same time
 +          * If using objective focus to make crisper pharynx after lawn imaging then the camera MOVES and the lawn is out of place
 +          * Possible to use same filter (YFP) for taking lawn picture and pharynx recording WITHOUT moving the camera up/down or focusing the objective using these settings respectively: ​
 +            * {{ :​wiki:​documentation:​wiki:​documentation:​nicolina:​image_from_ios_2_.jpg?​linkonly |}}
 +            * {{ :​wiki:​documentation:​wiki:​documentation:​nicolina:​image_from_ios_3_.jpg?​linkonly |}}
 +          * BUT ONLY IF:
 +            * The worms are already on the agar
 +            * The pharynx is focused on but the lawn remains out of focus
 +          * OTHERWISE you would then need to focus on the pharynx while worms are swimming in M9 and hope for the best because the focus changes once the worms hit the agar --> would need to focus using the objective again - transition between M9->agar without re-focusing already doesn'​t work well on the Zeiss
 +          * There should be a counterbalance for the optics on the other side of the stage - lifting the optics up and down is clunky and heavy - it's very difficult to hold the optics in the perfect place with one hand and screw the it into place with the other
 +      * Discussed with Luis about software to help automate the process
 +        - Automatic file saving and lawn photo taking to save time setting this up and waste less time once the first worm is in the field of view
 +        - Automatic re-focusing so there'​s no need to move the optics at all once it's in place and the lawn stays in the exact same position as in its photo
 +    * Due to these issues I only had time to record 1 video using the macroscope (0052) before giving up and switching to the Zeiss, by which point the worms were too starved and I had to starve new worms
 +      * 1xINF100 and 1xGRU101 video were taken with the Zeiss as a result and can be used for analysis
 +
  
wiki/documentation/wiki/documentation/nicolina/24.4.20.txt ยท Last modified: 2020/04/24 17:51 by nzjacic

Bitnami