Difference between revisions of "Optical Microscopy: Part 2 Report Outline"
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<li>Data</li> | <li>Data</li> | ||
<ul> | <ul> | ||
| − | <li>Include raw images of | + | <li>Include raw images of 0.84 um and 3.2 um fluorescent beads, the stained cell samples (with and without Cyto-D), the reference image paired with each sample image, and dark image paired with each sample image.</li> |
| − | + | ||
</ul> | </ul> | ||
<li>Analysis and Results</li> | <li>Analysis and Results</li> | ||
<ul> | <ul> | ||
| − | <li> | + | <li>For one reference, dark and sample image set, plot an intensity profile across the diagonals. Place all three profiles on a single set of axes. (Use the <tt>improfile</tt> command in MATLAB.)</li> |
<li>Use the reference and dark images to correct the raw image for nonuniform illumination and include the corrected image.</li> | <li>Use the reference and dark images to correct the raw image for nonuniform illumination and include the corrected image.</li> | ||
<li>For the bead samples on the one hand, and for the cell samples on the other hand, include histograms of the sample, reference, dark, and corrected images on a single plot of log(count) vs intensity. (You may plot the mere envelop of the histogram, for clarity purposes.) </li> | <li>For the bead samples on the one hand, and for the cell samples on the other hand, include histograms of the sample, reference, dark, and corrected images on a single plot of log(count) vs intensity. (You may plot the mere envelop of the histogram, for clarity purposes.) </li> | ||
Revision as of 20:21, 11 February 2015
- Update the apparatus section of your report to reflect the changes you made in part 2.
- Fluorescence imaging and flat-field correction
- Procedure
- Document the samples you used and how you captured images (camera settings, software used, etc…)
- Data
- Include raw images of 0.84 um and 3.2 um fluorescent beads, the stained cell samples (with and without Cyto-D), the reference image paired with each sample image, and dark image paired with each sample image.
- Analysis and Results
- For one reference, dark and sample image set, plot an intensity profile across the diagonals. Place all three profiles on a single set of axes. (Use the improfile command in MATLAB.)
- Use the reference and dark images to correct the raw image for nonuniform illumination and include the corrected image.
- For the bead samples on the one hand, and for the cell samples on the other hand, include histograms of the sample, reference, dark, and corrected images on a single plot of log(count) vs intensity. (You may plot the mere envelop of the histogram, for clarity purposes.)
- Document the steps you used to process the image.
- Discussion
- How did your beam expander design affect your images?
- Procedure
- Disrupting and imaging the actin network
- Procedure
- Document sample preparation, including difficulties, and recording conditions (particularly exposure time and gain selected in the camera settings)
- Data
- Include images of mouse embryonic fibroblasts + and - CytoD.
- Analysis and Results
- What contrast do you unveil between the + and - CytoD conditions?
- Discussion
- Can you quantify the morphological and physiological effect of CytoD treatment? What parameters would you choose to measure and report?
- Procedure
