| |
Resolvable Detail
The smallest resolvable detail imaged by an objective lens can be
estimated by 0.550µm/2xNumerical Aperture. That detail appears in the virtual
image produced by the objective lens, enlarged by the magnification of the lens.
The detail in the virtual image is projected via a tube lens onto the surface of
a CCD chip in a color camera. The smallest resolvable detail in the virtual
image observable with the camera is determined by the pixel periodicity of the
CCD chip. The output of the camera goes through a video controller board for
pixel interpolation and is transferred to the monitor for display. The smallest
resolvable detail observable at the monitor is determined by the monitor pixel
periodicity which is usually based on the display mode. In very high resolution
systems observable resolution is limited by the Red-Green-Blue triad periodicity
of the cathode ray tube.
The diagram below shows an example of the smallest resolvable detail at each
level in an imaging path from an objective lens to a monitor. The
resolution-limiting component is the one that has the largest detail size shown
for that objective lens.
| Monitor
Pixel Size |
Camera
Pixel Size |
VGA
(640x480) |
Ideal
(1600x1200) |
at
|
|
0.48mm |
0.25mm |
0.37
x 0.43mm |
|
| Objective |
Lens
Resolvable |
at
Virtual |
at
Analog Camera |
at
Monitor |
| Lens |
Detail |
Image |
1
CCD |
3 CCD |
(640x480) |
|
5x |
1.83µm |
9.15µm |
5.80µm |
3.32µm |
4.42µm |
| 10x |
0.92µm |
9.20µm |
2.90µm |
1.66µm |
2.21µm |
| 20x |
0.60µm |
12.0
µm |
1.45µm |
0.83µm |
1.11µm |
| 50x |
0.34µm |
17.0
µm |
0.58µm |
0.33µm |
0.44µm |
| 100x |
0.29µm |
29.0
µm |
0.29µm |
0.17µm |
0.22µm |
|
Field-of-View
The field-of-view for a microscope with oculars is equal to the
Ocular Field # / Objective Lens Magnification (if there are no magnifying
intermediate lenses). For Field # 20 oculars, the field-of-view is shown below,
along with the area displayed on a typical electronic imaging system.
| Obj |
Field-of-View
with Oculars |
Video |
| Lens |
Total
Diameter |
Perceived
Area |
Displayed
Area |
| 5x |
4.0 mm |
2.40 x
2.00 mm |
1.20 x
0.90 mm |
| 10x |
2.0 mm |
1.20 x
1.00 mm |
0.60 x
0.45 mm |
| 20x |
1.0 mm |
0.60 x
0.50 mm |
0.30 x
0.22 mm |
| 50x |
0.4 mm |
0.24 x
0.20 mm |
0.12 x
0.09 mm |
| 100x |
0.2 mm |
0.12 x
0.10 mm |
0.06 x
0.045mm |
When performing rapid screening through oculars, the area that information
is gleaned from is not the total area illuminated1,2. The
perceived area shown in the table is an average (some see more, while
others see less). But the reality is that the best one can expect to glean
information from is 75% of the diameter on the horizontal axis and 66% on
the vertical axis. The lower limit is 50%/33%. The Perceived Area data
reflects the average. Since a video image is 100% perceived, to achieve
the same screening area, a 2x lower objective is required. |

Total Diameter/Perceived Area |
- "How Did I Miss Those Cells?" Gary W. Gill, CT (ASCP), ADVANCE
for Medical Laboratory Professionals, Nov. 20, 1995
- "Visual Conspicuity, visual search and fixation tendencies of the
eye" Engel F.L., Vision Res. 17:95-108, 1977
|
|