User:Brian P. Josey/Notebook/2010/06/08: Difference between revisions

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I'm going over the basics of making an emulsion for the salad dressing project today.
I'm going over the basics of making an emulsion for the salad dressing project today.


[[User:Brian P. Josey|Brian P. Josey]] 15:18, 8 June 2010 (EDT) editing as I go, saving to protect. Final, clear write-up later. The scattered writing is to just job my memory, take quick notes and not get hung up on any of the details.
* use Tween 20
* quantum dots in water
* water with Tween in oil
* sonicate and see what happens
* quantum dot prep:
** green is more concentrated
** centrifuge with counter weight 5000 g for 3 min
** dilute 1:100 with fixing buffer
What I did:
Q dots:
* centrifuged as above
* 99 uL of buffer
* 1 uL of Q dots
Emmulsion:
mixed in this order
* 20 uL of Qdots
* 2 uL of Tween 20
* 78 uL of Mineral Oil
Observed: water is denser than the oil, created small surface at bottom of the tube, oil has greater cohesion, or tween pushes it down much more.
made flow cell: standard procedure
sonicated
observered: white milkiness as a direct result of sonication, more opaque on bottom half than on top half
flowed into flow cell, using two 10 uL pulls from the center of the tube, aiming to get a 50-50 mix of upper and lower halves. Observered white textured, thicker in some spots than in others.
Results from microscope: too thick, milky, poorly made flow cell and quantum dots are either everywhere, or some how got into the oil because: '''EVERYTHING IN THE SLIDE IS FLOURESCING GREEN!''' so use less quantum dots, higher oil concentration


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Emulsions

I'm going over the basics of making an emulsion for the salad dressing project today.

Brian P. Josey 15:18, 8 June 2010 (EDT) editing as I go, saving to protect. Final, clear write-up later. The scattered writing is to just job my memory, take quick notes and not get hung up on any of the details.

  • use Tween 20
  • quantum dots in water
  • water with Tween in oil
  • sonicate and see what happens
  • quantum dot prep:
    • green is more concentrated
    • centrifuge with counter weight 5000 g for 3 min
    • dilute 1:100 with fixing buffer


What I did: Q dots:

  • centrifuged as above
  • 99 uL of buffer
  • 1 uL of Q dots

Emmulsion: mixed in this order

  • 20 uL of Qdots
  • 2 uL of Tween 20
  • 78 uL of Mineral Oil

Observed: water is denser than the oil, created small surface at bottom of the tube, oil has greater cohesion, or tween pushes it down much more.

made flow cell: standard procedure

sonicated

observered: white milkiness as a direct result of sonication, more opaque on bottom half than on top half

flowed into flow cell, using two 10 uL pulls from the center of the tube, aiming to get a 50-50 mix of upper and lower halves. Observered white textured, thicker in some spots than in others.


Results from microscope: too thick, milky, poorly made flow cell and quantum dots are either everywhere, or some how got into the oil because: EVERYTHING IN THE SLIDE IS FLOURESCING GREEN! so use less quantum dots, higher oil concentration

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