Cluzel Lab Notebook
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Glycerol Stocks for MG1655 strains
Forgot to take cultures out on Friday, so doing this again.
Retry molds again
Will pour 2 molds using silicon chips to generate features, so it will be easier to focus. This must done in the humidity and temp. controlled room so features generate nicely. I am worried about desiccation of the agarose, so I'm going to keep it in the Humidity and Temp controlled room (HTC room) for as long as possible, and get the imaging equipment ready to see if the gel still distorts like on Wednesday. Jeff thinks that using the cover slip on top might not be necessary, but as long as it is placed on top carefully it should not 'squish' the features and it will help reduce moisture loss once the device is out of a high-humidity environment by limiting exposed surface area. There will be no exposed surface area in the actual device, so this hopefully will not be a problem.
Put cells on glass slide right before agarose is done setting in normal conditions, move into HTC room and remove from mold and put on top of cells immediately. Cover with square cover slip to maintain moisture.
Check spot size before agarose application
Spotted 5 0.2 uL dots of stationary phase cells onto a glass slide, imaged to check spot diameter. On the order of 0.5 to 1.5 CCD chips in spot diameter, if correctly applied. Spots dry within 1 minute. Salt crystals formed in dried spots. (Images saved in C:\MM\images\dbg\expt* dir on camera computer)
Compare spot size before and after agarose application
Prep cells to remove salt crystalization and dilute
Spotting on slide
Cells remain on agarose, with the majority constrained to one region (little or no 'wicking' observed). (Images saved in C:\MM\images\dbg\expt3* dir on camera computer)
Transfer from slide to slide
Do the cells transfer well? Is there a clear transfer boundary?