scale bar with the leica
DESCRIPTION
scaleTRANSCRIPT
Adding a scale bar to images taken with the Leica Fluorescent Microscope
Adding a Scale Bar to Images Taken with the Leica Fluorescent Microscope
Using Built in Software (Leica Application Suite) more convenient for a couple images
Take an image Aquire tab -> Acquire Image
Switch to Process tab
Select your image from the dashboard at the bottom of the screen
Switch to Calib. Sub-tab
Click Manual (Draw Line)
Enter the pixels and length for the objective you used according to the chart below:
Lens MagnificationPixelsLengthUnit of Length
Leica 5x107200m
Leica 10x107100m
Leica 20x438200m
Leica 40x431100m
Leica 63x680100m
Select microns as the unit
Click Apply
Switch to Annotate sub-tab
In the Scale Bar tab, click Show
You can now click and drag the scale bar to place it wherever you please
You may also change the length by selecting User Length and typing a new value
Format the font and colors in the font box
In the Actions box, click Merge
Right click on the large image, click Save As
Save the image with scale bar to you folder.
Using ImageJ Easier for many images
Save all images with the magnification in the name or on the folder!!! (ex: 5x, 10x)
Download ImageJ: http://rsb.info.nih.gov/ij/download.htmlInstall ImageJ
Open the first picture file into ImageJ: ctrl+O (or) file -> open
Set Scale: Analyze -> Set Scale
Enter pixel and distance values and unit of length according to table below:
Lens MagnificationDistance in PixelsKnown DistanceUnit of Length
Leica 5x107200m
Leica 10x107100m
Leica 20x438200m
Leica 40x431100m
Leica 63x680100m
Check the Global length box!
Click: Analyze -> Tools-> Scale Bar
Enter in the desired length of scale bar in the Width in box
Format the bar with the following boxes
You can now quickly add scale bars to your other pictures:
Open image
Click: Analyze -> Tools-> Scale Bar
Format scale bar
If you clicked the global length box, you will not have to set the scale for each image until you edit an image that used a different objective.
If you do not know the Distance in pixel or known distance
Get a measuring standard slide (or mm ruler for the dissecting microscopes).
Take a picture, record the objective magnification as well as any projector lenses between the objective and camera.
Open picture in ImageJ
Zoom in on the standardization lines with the magnifying glass tool (10th icon on tool bar)
Use the line tool (5th icon on tool bar) and draw a line along the known length of the standardization lines (hint: hold shift to constrain the line to right angles)
Go to Set Scale: Analyze -> Set Scale
The value in distance in pixels is your Distance in Pixels and the standardization length is the known distance
Additional Pixel/Length Tables (Note: there are multiple lenses! The projector lens sits in the collar adapter right beneath the camera.)
Olympus Compound Microscope
lens 1 objectivelens 2 projectorDistance in PixelsKnown Distance (m)
42.5750.00666.67
43.3829.68550.00
45826.04366.67
102.5791.01283.33
103.3748.03200.00
105839.06150.00
202.5742.00133.33
203.3746.10100.00
205742.0266.67
402.5783.0070.00
403.3801.0953.33
405749.1533.33
1002.5757.0026.67
1003.3765.0220.00
1005742.0013.33
WILD Dissecting Microscope
lens 1 screw onlens 2 bottomlens 3 objectivelens 4 projectorDistance in PixelsKnown Distance (m)
0.5162.52.1825.00
0.5163.32.3020.00
0.51651.7310.00
0.51122.51.7610.00
0.51123.32.2810.00
0.511252.457.00
0.51252.52.146.00
0.51253.31.944.00
0.512552.123.00
0.51502.52.163.00
0.51503.31.932.00
0.515051.461.00
none162.50.342.00
none163.30.452.00
none1650.682.00
none1122.50.682.00
none1123.30.912.00
none11251.372.00
none1252.51.532.20
none1253.31.761.90
none12551.681.20
none1502.51.811.30
none1503.31.861.00
none15051.680.60