User:Brian P. Josey/Notebook/2010/06/23

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1:200 Emulsion

Unsatisfied with yesterday's very dense results, I diluted the emulsion with an additional 90 μL of mineral oil. With the original volume of 90 μL of emulsion, 10 μL were drawn off for a flow cell, this brings the emulsion down to 1 part ferritin solution to 200 parts mineral oil. I am going to film this one also over a fifteen minute stretch, and see if I can get a better picture of any movements.

Ferritin Results and Tracking

I put a flow cell of the above emulsion on the microscope and mounted the magnet on the stage. Like I did yesterday, I filmed it a 1 fps for fifteen minutes, and compiled the images into a movie. Here is the video sped up to 10 fps in 90 seconds:

<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/vap3CVOqVPc&hl=en&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/vap3CVOqVPc&hl=en&fs=1" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object> </html>

Here I had the magnet at the top of the image, which is weird considering that the it appears the droplets are moving to the lower left corner. As Koch suggested in the talk page, it would make sense that the objective has been magnetized and is having a significant effect on the proteins. To take a closer look at what was happening, I sped up the film even further, up to 60x real time. Here the whole fifteen minutes passes in fifteen seconds:

<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/rOJOphfRdgk&hl=en&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/rOJOphfRdgk&hl=en&fs=1" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object> </html>

Here it is very clear that there is a general trend in the droplets to move to the lower left corner of the image. I was then able to match up some droplets in the first frame with the last frame. Here, the numbers represent individual droplets, and the colors represent how long I was able to track them. Blue is for the droplets I could follow through the whole clip, while red is for those that disappear by going out of focus or leaving the field of view. Here is the first, left, and last, right, frame from the film with the droplets numbered:

10 Initial Numbers.png 10 Final Numbers.png

From this and carefully watching the footage, I was able to create a third image with arrow representing the general path each droplet took:

10 Arrows.png

Of course, the droplets move around in a more complicated way than I've indicated when you account for Brownian motion and other random noise. But it is clear that they are all moving in one direction, so they are under the influence of a force of some kind, which is probably magnetic. Because it is moving away from the magnet however, it is pretty clear that any residual magnetization on the objective is having a pronounced effect. I was then able to extend this further, and traced the general paths for twenty five droplets:

25 Arrows.png

The yellow arrow here represents a droplet that I think merged into another one. Steve Koch 22:44, 23 June 2010 (EDT): Maybe just popped into another focal plane? Hard to say. Nice work with analysis, must have taken a long time!