Acquire Images
Objective List
All require 0.17mm coverslip unless noted
2 5x Plan-Neofluar
1
NA . . . . . . . . . . . . . . . . . 0.075
WD(mm) . . . . . . . . . . . . . . . 9.5
Depth of Field (560nm) . . . . . . 139
m
m
Part # . . . . . . . . . . . . . . . 440310
5x Plan-Neofluar
1
NA . . . . . . . . . . . . . . . . . 0.075
WD(mm) . . . . . . . . . . . . . . . 9.5
Depth of Field (560nm) . . . . . . 31.1
m
m
Part # . . . . . . . . . . . . . . . 440310
10x Plan-Neofluar
1
NA . . . . . . . . . . . . . . . . . 0.30
WD(mm) . . . . . . . . . . . . . . . 5.6
Depth of Field (560nm) . . . . . . 8.69
m
m
Phase . . . . . . . . . . . . . . . . Ph-1
DIC . . . . . . . . . . . . . . . . . DIC II
Part # . . . . . . . . . . . . . . . 440331
20x Plan-Apochromat
3
NA . . . . . . . . . . . . . . . . . 0.80
WD(mm) . . . . . . . . . . . . . . 0.55
Depth of Field (560nm) . . . . . . 1.34
m
m
DIC . . . . . . . . . . . . . . . . . DIC II
Part # . . . . . . . . . . . . . . . 440640
40x OIL Plan-Apochromat
3
NA . . . . . . . . . . . . . . . . . . 1.4
WD(mm) . . . . . . . . . . . . . . . .19
Depth of Field (560nm) . . . . . . 514nm
DIC . . . . . . . . . . . . . . . . .DIC III
Part # . . . . . . . . . . . . . . . 440865
50x EC Epiplan Air EC-Epiplan
4
NA . . . . . . . . . . . . . . . . . . 0.7
WD(mm) . . . . . . . . . . . . . . . .1.1
Depth of Field (560nm) . . . . . . 1.27
m
m
Coverslip . . . . . . . . . None required
DIC . . . . . . . . . . . . . . . . .DIC III
Part # . . . . . . . . . . . . 422070-9960
63x OIL Plan-Apochromat
3
NA . . . . . . . . . . . . . . . . . . 1.4
WD(mm) . . . . . . . . . . . . . . .0.19
Depth of Field (560nm) . . . . . . 561nm
DIC . . . . . . . . . . . . . . . . .DIC III
Part # . . . . . . . . . . . . . . . 440762
100x OIL Plan-Apochromat
3
NA . . . . . . . . . . . . . . . . . . 1.4
WD(mm) . . . . . . . . . . . . . . .0.17
Depth of Field (560nm) . . . . . . 514nm
DIC . . . . . . . . . . . . . . . . .DIC III
Part # . . . . . . . . . . . . . . . 440780
Objective Notes:
1 ) Plan-Neofluar: very good quality-correct-
ed for spherical aberration and corrected
for chromatic aberration in red and blue
wavelengths
2 ) Achroplan: good quality-corrected for
adequate field flatness (spherical aberra-
tion) and chromatic aberration in red and
blue wavelengths to a lesser degree than the
Neofluar A compromise for special applica-
tions like long working distance
3 ) Plan-Apochromat: excellent quality cor-
rect for spherical aberration and corrected
for chromatic aberration in red, blue and
green wavelengths
4 ) EC Epiplan: Enhanced Contrast Univer-
sal objectives for materials microscopy
allowing brightfield, differential interfer-
ence contrast and polarization methods
HD versions provide darkfield capability in
addition
Acquire Image Stacks (Z-Series).
1. Select Imaging > Acquire Image Stack
Spatial Calibration:
Olympus Microfire color CCD
2.5x Bin 1 . . . . . . . . . . 2.8
m
m/pixel
5x Bin 1 . . . . . . . . . . . 1.45
m
m/pixel
10x Bin 1 . . . . . . . . . . 0.73
m
m/pixel
20x Bin 1 . . . . . . . . . . 0.36
m
m/pixel
40x Bin 1 . . . . . . . . . . 0.18
m
m/pixel
50x Bin 1 . . . . . . . . . . 0.14
m
m/pixe
63x Bin 1 . . . . . . . . . . 0.11
m
m/pixel
100x Bin 1 . . . . . . . . . . 0.07
m
m/pixel
Original images found in: \stereology work-
space\Tutorials\Calibration Images
2. Select Options... then Set top and
bottom, this will change the appearance
of the Image acquisition menu allow-
ing selection of a focus point above the
sample and a focus point below. Set the
top of the stack and the bottom of the
stack.
3. Set the distance between images to
any reasonable number from .1 mm to
With StereoInvestigator
2mm depending on the objective.
4. Set the delay between images to 750
ms, this will allow the stage time to settle
before the next image is acquired.
5. Acquire image stack.
Multichannel acquire with bring up ad-
vanced options for fluorescence imaging.
6. Save image stack.
Setting delay to at least 750ms will re-
duce stage chatter during image acquisi-
tion.
