Daniel:Notebook/HiResChrPaint/2014-9-23: Difference between revisions
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===DMD-488 and Cot1-546=== | ===DMD-488 and Cot1-546=== | ||
<gallery widths=500px heights=500px perrow=3 caption="Image series for DMD-488 and Cot1-546"> | Note that all thresholds are set the same in this series as well. They are {min,max}: Bright Field {50,200}; 488 {40,60}; 546 {20,80}. | ||
<gallery widths=500px heights=500px perrow=3 caption="Image series for DMD-488 (green) and Cot1-546 (red)"> | |||
File:20140923_DMD488_Cot1546.lif_Series_2048pxSnapshot.jpg|2048 pixels (1) | File:20140923_DMD488_Cot1546.lif_Series_2048pxSnapshot.jpg|2048 pixels (1) | ||
File:20140923_DMD488_Cot1546.lif_Series_2048px_2Snapshot.jpg|2048 pixels (2) | File:20140923_DMD488_Cot1546.lif_Series_2048px_2Snapshot.jpg|2048 pixels (2) | ||
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</gallery> | </gallery> | ||
====Comparison==== | |||
The following gallery is the comparison between the 2048 pixel images, 1,2 and 6, showing the degradation between the image series. The '''Difference''' method subtracts the two pixel images (image 2 or 6 subtracted from image 1) and the '''Minimum''' method chooses the lower of the two pixel values and displays that in the comparison. Using the difference method, spots that are those that are different between the two images. Using the minimum method, pixels that show up are those that are the same. All images were thresholded (as above) and viewed as maximum projection prior to image comparison. | |||
<gallery widths=600px heights=600px perrow=2 caption="Image comparisons for 2048 pixel images, DMD-488 (green) and Cot1-546 (red)" > | |||
File:20140923_DMD488_Cot1546.lif_Series_2048px_1-2Combined_DifferenceSnapshot1.jpg|1-2 Comparison: Difference | |||
File:20140923_DMD488_Cot1546.lif_Series_2048px_1-2Combined_MinimumSnapshot1.jpg|1-2 Comparison: Minimum | |||
File:20140923_DMD488_Cot1546.lif_Series_2048px_1-6Combined_DifferenceSnapshot1.jpg|1-6 Comparison: Difference | |||
File:20140923_DMD488_Cot1546.lif_Series_2048px_1-6Combined_MinimumSnapshot1.jpg|1-6 Comparison: Minimum | |||
</gallery> | |||
===Padlock Probes-488 Cot1-546=== | |||
Note that all thresholds are set the same in this series as well. They are {min,max}: Bright Field {50,200}; 488 {40,70}; 546 {40,70}. | |||
<gallery widths=500px heights=500px perrow=3 caption="Image series for PPRL-488 and Cot1-546"> | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_1Snapshot_All1.jpg|2048 pixels (1) | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_2Snapshot_All1.jpg|2048 pixels (2) | |||
File:20140923_PPRL488_Cot1546.lif_Series_1024px_3Snapshot_All1.jpg|1024 pixels (3) | |||
File:20140923_PPRL488_Cot1546.lif_Series_512px_4Snapshot_All1.jpg|512 pixels (4) | |||
File:20140923_PPRL488_Cot1546.lif_Series_256px_5Snapshot_All1.jpg|256 pixels (5) | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_6Snapshot_All2.jpg|2048 pixels (6) | |||
</gallery> | |||
====Comparison==== | |||
The following gallery is the comparison between the 2048 pixel images, 1,2 and 6, showing the degradation between the image series. The '''Difference''' method subtracts the two pixel images (image 2 or 6 subtracted from image 1) and the '''Minimum''' method chooses the lower of the two pixel values and displays that in the comparison. Using the difference method, spots that are those that are different between the two images. Using the minimum method, pixels that show up are those that are the same. All images were thresholded (as above) and viewed as maximum projection prior to image comparison. | |||
<gallery widths=600px heights=600px perrow=2 caption="Image comparisons for 2048 pixel images, DMD-488 (green) and Cot1-546 (red)" > | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_1-2Combined_DifferenceSnapshot_All1.jpg|1-2 Comparison: Difference | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_1-2Combined_MinimumSnapshot1.jpg|1-2 Comparison: Minimum | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_1-6Combined_DifferenceSnapshot1.jpg|1-6 Comparison: Difference | |||
File:20140923_PPRL488_Cot1546.lif_Series_2048px_1-6Combined_MinimumSnapshot1.jpg|1-6 Comparison: Minimum | |||
</gallery> | |||
[[Category:HRCP]] [[Category:Microscopy]] [[Category:20140919]] | [[Category:HRCP]] [[Category:Microscopy]] [[Category:20140919]] |
Latest revision as of 21:30, 25 September 2014
Slide FISH (Started 09/19/2014)[edit]
Microscopy[edit]
For today's microscopy, I tried something new in order to help determine some of the properties of the microscope/sample. I took an image series for both the DMD and the PPRL samples using different resolutions in the following order (values are for one side, total image is number of pixels squared): 2048 pixels, 2048 pixels, 1024 pixels, 512 pixels, 256 pixels, 2048 pixels. All other properties, gain, x,y,z locations, etc. were kept constant. This will allow me to see (a) if the resolution of the image seems to matter and (b) fluorophore quenching from multiple rounds of imaging.
DMD-488 and Cot1-546[edit]
Note that all thresholds are set the same in this series as well. They are {min,max}: Bright Field {50,200}; 488 {40,60}; 546 {20,80}.
- 20140923 DMD488 Cot1546.lif Series 2048pxSnapshot.jpg
2048 pixels (1)
- 20140923 DMD488 Cot1546.lif Series 2048px 2Snapshot.jpg
2048 pixels (2)
- 20140923 DMD488 Cot1546.lif Series 1024px 3Snapshot.jpg
1024 pixels (3)
- 20140923 DMD488 Cot1546.lif Series 512x 4Snapshot.jpg
512 pixels (4)
- 20140923 DMD488 Cot1546.lif Series 256px 5Snapshot.jpg
256 pixels (5)
- 20140923 DMD488 Cot1546.lif Series 2048px 6Snapshot.jpg
2048 pixels (6)
Comparison[edit]
The following gallery is the comparison between the 2048 pixel images, 1,2 and 6, showing the degradation between the image series. The Difference method subtracts the two pixel images (image 2 or 6 subtracted from image 1) and the Minimum method chooses the lower of the two pixel values and displays that in the comparison. Using the difference method, spots that are those that are different between the two images. Using the minimum method, pixels that show up are those that are the same. All images were thresholded (as above) and viewed as maximum projection prior to image comparison.
- 20140923 DMD488 Cot1546.lif Series 2048px 1-2Combined DifferenceSnapshot1.jpg
1-2 Comparison: Difference
- 20140923 DMD488 Cot1546.lif Series 2048px 1-2Combined MinimumSnapshot1.jpg
1-2 Comparison: Minimum
- 20140923 DMD488 Cot1546.lif Series 2048px 1-6Combined DifferenceSnapshot1.jpg
1-6 Comparison: Difference
- 20140923 DMD488 Cot1546.lif Series 2048px 1-6Combined MinimumSnapshot1.jpg
1-6 Comparison: Minimum
Padlock Probes-488 Cot1-546[edit]
Note that all thresholds are set the same in this series as well. They are {min,max}: Bright Field {50,200}; 488 {40,70}; 546 {40,70}.
- 20140923 PPRL488 Cot1546.lif Series 2048px 1Snapshot All1.jpg
2048 pixels (1)
- 20140923 PPRL488 Cot1546.lif Series 2048px 2Snapshot All1.jpg
2048 pixels (2)
- 20140923 PPRL488 Cot1546.lif Series 1024px 3Snapshot All1.jpg
1024 pixels (3)
- 20140923 PPRL488 Cot1546.lif Series 512px 4Snapshot All1.jpg
512 pixels (4)
- 20140923 PPRL488 Cot1546.lif Series 256px 5Snapshot All1.jpg
256 pixels (5)
- 20140923 PPRL488 Cot1546.lif Series 2048px 6Snapshot All2.jpg
2048 pixels (6)
Comparison[edit]
The following gallery is the comparison between the 2048 pixel images, 1,2 and 6, showing the degradation between the image series. The Difference method subtracts the two pixel images (image 2 or 6 subtracted from image 1) and the Minimum method chooses the lower of the two pixel values and displays that in the comparison. Using the difference method, spots that are those that are different between the two images. Using the minimum method, pixels that show up are those that are the same. All images were thresholded (as above) and viewed as maximum projection prior to image comparison.
- 20140923 PPRL488 Cot1546.lif Series 2048px 1-2Combined DifferenceSnapshot All1.jpg
1-2 Comparison: Difference
- 20140923 PPRL488 Cot1546.lif Series 2048px 1-2Combined MinimumSnapshot1.jpg
1-2 Comparison: Minimum
- 20140923 PPRL488 Cot1546.lif Series 2048px 1-6Combined DifferenceSnapshot1.jpg
1-6 Comparison: Difference
- 20140923 PPRL488 Cot1546.lif Series 2048px 1-6Combined MinimumSnapshot1.jpg
1-6 Comparison: Minimum