Lab 1 Report -- Nathan S Lachenmyer

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Calibration

All calibrations and experiments done at 20 mW.

Position Calibration

  • Learned to make microfluidic devices out of double-stick tape and a slide -- this was really neat!
  • Made two samples with 1 um microspheres
    • One sample with free-floating spheres (in H2O)
    • Another one with 'stuck' spheres in NaCl (I cheated :( )
  • Took a position calibration on the optical trap setup, seen below

1um cal.png

calibration: 502463 Volts / m = 1.99 um / volt

Trap Stiffness Calibration

  • Did all three versions of the trap calibration, resulting in the following trap spring constants:
Method Trap Stiffness (pN / nm)
Stokes 2.26e-5 pN/nm
Equipartition Theorem 1.23e-5 pN/nm
PSD *Need Bandwidth of DAQ*

Experiments

E Coli

  • Learned to culture E Coli (sort of)
  • Examined the various cultures to determine which ones had the fastest spinners / spinners in the appropriate direction with Steve
  • E Coli weren't spinning very well -- swapped out the blue LED for a Red LED to determine if the wavelength made a difference
    • As far as we could tell, the LED color made no difference
    • We also couldn't figure out why the E Coli were spinning so slowly
  • Worked with Steve to cut the flagella of E Coli by drawing them in and out of a pipette multiple times
    • This improved the spinning frequency of the E Coli!

Ecoli.png

Doing an FFT analysis of the QPD spectrum, I find that the low-frequency components are pretty noisy (unfortunately). However, there is a small peak at approximately 6 Hz -- according to the appleyard paper, the e coli flagellar motor spin frequency is approximately 4-10 Hz, so this is in good agreement.

DNA Tethers

  • Learned to make the DNA tethers
  • Was able to trap a microsphere tethered to the slide via DNA

Dna raw.png

Stretchiness of the DNA: