2005-06-05

(2.14 pm)
Plan: 
- get an image using the cheap CCD and the halogen study lamp as the light source
- in the other words, repeat previous tests for picture purposes
- get an image of 1um bead using the cheap CCD

- the cheap camera is useful to debug the microscope, that way one would not have to worry about damaging the HAMAMATSU camera by inadvertently exposing it to too much light.
Things that I did
- the electron multiplier CCD camera was replaced with the cheap CCD camera (see Camera.jpg)
- the camera was connected to the TV
- remove the mirror to create space for the bright light source.  The light source was the white study lamp that Bernie purchased (see setup1-off.jpg and setup1-on.jpg)

(2.25 pm)
An image was observed on the TV screen (see Tv1.jpg)
The image of what???
- rotate the CCD, the image was rorated  --> the source is not a dusty CCD  (see Tv2.jpg and Tv3.jpg)
- the upper and/or objectives was rotated, the image was not rotated --> the source is not a dusty upper and/or objective 

(5.00 pm)
- the old lower objective were taken out --> a brighter illumination (as seen from the illumination spot above the upper objective)  (see no-lower-objective.jpg)
- the bright and unfocused illumianation should be bad due to the larger area of photobleached region
- could not see image

- the old lower objective (not the TIRF objective) was installed again --> much tighter illumination spot than without lower objective was obtained, as expected
- the image of the 1um dye (2/100 dillution factor) was observed clearly (the video file: 1um-bead.3gp).  The image was not optimezed.

- it seems to me that the problem that the allignment change as we focus the upper objective
- trick: 
  1. open the box above the upper objective to allign the image
  2. use maximum intensity, if the image hits the CCD, the screen will be brighter.  If you don't see much, focus the upper objective

(6.00 pm)
- clean up

The pictures and videos are stored at http://www.dna.caltech.edu/~hariadi/spFRET/2005-06-05/