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Chapter 5: Analysis Tools
5-63
the spot the software will define. The greater the sensitivity number the more of the
object the software will find.
**HINT: Selecting a brighter pixel within the band (or spot) will have magic wand draw
a tighter area of interest. Correspondingly, a less intense pixel will draw a larger
perimeter around the selected spot**
Spot Type
Under the Spot Type heading the Bright Spots selection should be checked if the image
contains bright spots on a dark background (e.g. ethidium bromide stained fluorescent
gels and chemiluminescent blots). The Dark Spots selection should be made when the
sample contains dark sample on a light background (film, coomassie blue protein gels
etc..). The software should automatically determine this selection, but the user may
have to override it if the software incorrectly evaluates an image. Remember as in all
portions of AlphaEaseFC software if the image has been reversed (negative image)using
the Reverse selection on the Contrast Adjustment window the Dark Spot selection
should not be made. Reverse is a visual alteration only and does not effect the original
image capture data. (EG a chemiluminescent blot is captured using the Reverse mode in
order for it to appear as film, the correct selection is still Bright Spots).
Outline Type
Border Outline will use the edge detection methods to determine the spot area. The Box
Outline will draw a box shape around the sample usually including a significant portion
of background. User preference and image particulars will determine which outline type
is best.
Previous Location
Previous Location will automatically select the spot on the image last selected for magic
wand and apply any changes made for sensitivity. This feature is designed to quickly
evaluate how the different sensitivity numbers will affect the drawn area using the same
selected point in a spot. The X,Y coordinates are given for the previous spot as well.
Exit
Exit will remove the Magic Wand Parameter Window.
Auto Spot:
Auto Spot is a feature designed to find multiple spots within a region of interest. It is
accessed through AutoSpot under Auto Detect in the Spot Denso tab. Once it is selected
the following tab will appear and the cursor will be ready to draw an area of interest.
- AlphaImager™ IS-2200 1
- For Windows 2000/XP 1
- AlphaImager 2
- 2200 – 2
- AlphaEaseFC User’s Manual 2
- 1.2.1 System Components 8
- 1.2.2 System Placement 8
- 1.2.3 Cable Connections 8
- 1.5 System Information 15
- 1.6 Acquiring an Image 16
- 1.6.1 Focusing on an Image 17
- Selecting a Filter 21
- Displays 23
- Imaging Functions 32
- 2.1 Contrast Adjustment 33
- Black Level Adjustment 34
- White Level Adjustment 35
- Gamma Setting Adjustment 35
- The Auto Contrast Selection 36
- The Reverse Button 37
- The Grid Button 37
- 2.2 Tool Bar 40
- Sample Printouts 42
- 2.3 Tool Box 43
- 2.4 Status Bar 43
- Chapter 3: Drop Down Menus 46
- File Open 47
- File Overlay 48
- File Close 48
- File Save and Save As 49
- File Types: 50
- Chapter 3: Drop Down Menus 52
- 3.2 The Edit Menu 53
- Reset and Clear 54
- 3.3 The Image Menu 55
- Conversion 56
- Flat Field Calibrate 57
- 3.4 The Setup Menu 59
- Load Defaults 60
- Preferences 63
- 3.5 The Overlay Menu 66
- 3.6 The Utilities Menu 69
- Notepad 70
- 3.7 The View Menu 71
- Zoom Functions 72
- Show Annotations 72
- 3.8 The Help Menu 73
- Chapter 4: The Image 76
- 4.2 The Rotate / Flip Tool 77
- 4.3 Histogram 78
- 4.4 Automatic Enhancement 79
- 4.5 Annotations 81
- Chapter 4: Enhancement Tools 82
- Text Background and Font 83
- Text Angles 84
- The Drawing Tools 84
- The Editing Tools 86
- 4.6 False Color 88
- Palettes 2 through 11 89
- 4.7 Image Filters 90
- Sharpening Filters 92
- 3-D (Contour) Filters 92
- Noise Filters 93
- Smoothing Filters 93
- Edge Filters 93
- Horizontal Edge Filter 93
- Vertical Edge Filter 94
- Custom Filter 94
- The UNDO Button 94
- Examples of Filter Results 94
- 4.8 Movie Mode 95
- Frame Stacking 98
- BLANK PAGE 100
- Tools in the Toolbox 102
- Setting up an ARRAY Template 103
- Aligning the Template 104
- Measuring Density 105
- The INVERT Box 106
- Outputting Data 106
- Output Dialog Box 107
- Introduction 108
- Chapter 5: Analysis Tools 110
- Manually Selecting Bands 111
- Automatic Band Finding 111
- Special Functions: 115
- Snap to Peak 115
- The INVERT Button 115
- Calculating R 116
- Values 116
- The Graph Tool 117
- Auto Grid 118
- Setting up the Lane Template 119
- Specifying the Scan Width 120
- Scanning the Image 121
- Graph Display 122
- The Active Graph 122
- The Data Window 123
- Peak Detection Algorithms 125
- Set parameters dialog box 129
- Reset Base 130
- Base Subtract 130
- Interpreting 1D-Multi Data 130
- V. Line 130
- 3-D View 131
- Smooth Data 131
- The Invert Button 132
- Overlaying Graphs 133
- 1D-Multi 134
- Band Scoring 139
- Outputting Quantitative Data 141
- Exporting Graphs 141
- Exiting 1D-Multi 142
- Auto Lane 142
- Black Bands or White Bands 142
- Sensitivity Adjustment Bar 143
- Area of Interest Drawing 143
- Output Data 146
- Baseline 147
- Options 147
- Editing Peak Boundaries 147
- Auto Lane Editing Features 148
- Manual Band Matching 156
- Band Matching Results 157
- 5.4 Spot Density Tools 161
- Magic Wand and AutoSpot 163
- Magic Wand: 163
- Magic Wand Sensitivity 163
- Area of Interest 165
- Bright Spot vs Dark Spot 166
- Find Spots 166
- Background Threshold 166
- Moving Objects 168
- Deleting Objects 168
- Saving and Loading Objects 168
- Object Libraries 169
- Spot Density Measurements: 169
- Data Definitions 170
- Inverting the Data 170
- No Background 171
- Automatic Background 171
- Manual Background 172
- The Standard Curve Toolbox 173
- Displaying the Curve 175
- Outputting Quantitation Data 176
- Manual Count Tools 177
- Auto Count Tools 177
- Manual Count 179
- Automatic Count 180
- Editing Tools 182
- Displaying Spots 183
- Using the ADD SPOT Tool 183
- Using the Erase Spot Tool 184
- Spot Count Data 184
- Outputting Auto Count Data 185
- 5.6 The Scoring Function 187
- Scoring the Sample 188
- 5.7 The Ruler Function 190
- Appendix A: Opening 192
- AlphaEaseFC Files 192
- Weight Library Files 194
- To Change the Security Setup 196
- Security Features Dialog Box 197
- Instructions 202
- Cabinet Setup 203
- Camera Installation 206
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