Changes between Version 2 and Version 3 of Research/LhARA/RadiationBiology/BMthdlgy/ImprvCllSrvvl


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Timestamp:
Apr 16, 2026, 5:44:00 PM (7 weeks ago)
Author:
ccd24
Comment:

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  • Research/LhARA/RadiationBiology/BMthdlgy/ImprvCllSrvvl

    v2 v3  
    22
    33- Reduce the irradiation time
    4  - Gain an accurate understanding of how long HeLa cells can withstand vertical
     4 1) Gain an accurate understanding of how long HeLa cells can withstand vertical
    55  - Current limits are that they can survive 20 minutes but not ~1.5 hours
    6  - Being aware of the issue
    7  - Reducing the time between shots from 20s to maybe 1s, definitely to 5s
    8  - Not irradiating RCF in the same carousel
     6 2) Being aware of the issue
     7 3) Reducing the time between shots from 20s to maybe 1s, definitely to 5s
     8 4) Not irradiating RCF in the same carousel
    99  - A combination of these two would bring the irradiation time from roughly 16 mins to roughly 4 mins
    1010- Dealing with potential causes: Temperature, CO2, lack of media, weak cell line
    11  - Heat the carousel in which the cells are placed
    12  - Keep the carousel rotating so that the cells are consistently wet
    13  - Change the cell dish lid so that more media can be added through a second hole
     11 5) Heat the carousel in which the cells are placed
     12  - Can this be done by placing the carousel in the incubator as well?
     13 6) Keep the carousel rotating so that the cells are consistently wet
     14 7) Change the cell dish lid so that more media can be added through a second hole
    1415  - Would also help regulate temperature
    15  - Change the cell line from HeLa to the super-resistant ones used in Marie's lab
     16 8) Change the cell line from HeLa to the super-resistant ones used in Marie's lab
    1617  - Requires testing the cells at Birmingham and repeating the experiments
    1718  - Not certain it will work
    1819- Equipment factors
    19  - Place a thermostat in the incubator to check that it is not too warm
     20 9) Place a thermostat in the incubator to check that it is not too warm
    2021  - Robbie has said he will build a monitor using a Raspberry Pi
    21  - Use Marie's lab to allow easier observation
     22 10) Use Marie's lab
    2223  - Need to inform Marie of when so that she can book in the incubator
     24  - Can do x-ray comparison here
     25  - Much bigger fume cupboards, so 2 people could work at once and reduce the load
     26  - Comet analysis
     27  - Easier day-to-day observation
     28
     29
     30Personal Thoughts
     31There are two things we do not know. The fastest we can irradiate cells and the time cells can survive vertically.
     32I think if we do 2, 3, and 4, we can get the time the cells are vertical for to less than 20 minutes. However, other factors may cause these to change, such as needing to take more shots, so I think it would be sensible to test how long the cells can last vertically (1) and see how much leeway we have. If there is no time, then I would go to SCAPA and test 5 and 6. If time allows, might as well test 7 and 8, but this feels like it will take longer for the new lids and the subsequent work of the new cell line.
     33
     34Either way, a dry run is completely necessary before more irradiations can be done.
     35
     36Finally, I would definitely install a thermostat in the incubator; a monitor from Robbie would be great, but essential.
     37
     38I do not know enough about whether to use Marie's lab.
     39If we do not, we need to acquire an inverted microscope and a multi-chamber haemocytometer???
     40
    2341
    2442