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
Unit of
Select microns as
Magnification Pixels Length
Length
the unit
Leica 5x
107
200
m
Click Apply
Leica 10x
107
100
m
Switch to
Leica 20x
438
200
m
Annotate sub-tab
Leica 40x
431
100
m
In the Scale Bar
Leica 63x
680
100
m
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.html
Install 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
Distance in
Known
Unit of
Magnification
Pixels
Distance
Length
Leica 5x
107
200
m
Leica 10x
107
100
m
Leica 20x
438
200
m
Leica 40x
431
100
m

Leica 63x
680
Check the Global length box!

100

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
lens 2
objective
projector
4
4
4
10
10
10
20
20
20
40
40
40
100
100
100

2.5
3.3
5
2.5
3.3
5
2.5
3.3
5
2.5
3.3
5
2.5
3.3
5

Distance in
Known Distance
Pixels
(m)
750.00
666.67
829.68
550.00
826.04
366.67
791.01
283.33
748.03
200.00
839.06
150.00
742.00
133.33
746.10
100.00
742.02
66.67
783.00
70.00
801.09
53.33
749.15
33.33
757.00
26.67
765.02
20.00
742.00
13.33

WILD Dissecting Microscope

lens 1
lens 2
screw on
bottom
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
none
none
none
none
none
none

1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1

lens 3
lens 4
Distance
objective projector in Pixels
6
2.5
2.18
6
3.3
2.30
6
5
1.73
12
2.5
1.76
12
3.3
2.28
12
5
2.45
25
2.5
2.14
25
3.3
1.94
25
5
2.12
50
2.5
2.16
50
3.3
1.93
50
5
1.46
6
2.5
0.34
6
3.3
0.45
6
5
0.68
12
2.5
0.68
12
3.3
0.91
12
5
1.37

Known
Distance
(m)
25.00
20.00
10.00
10.00
10.00
7.00
6.00
4.00
3.00
3.00
2.00
1.00
2.00
2.00
2.00
2.00
2.00
2.00

none
none
none
none
none
none

1
1
1
1
1
1

25
25
25
50
50
50

2.5
3.3
5
2.5
3.3
5

1.53
1.76
1.68
1.81
1.86
1.68

2.20
1.90
1.20
1.30
1.00
0.60