Beruflich Dokumente
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ZEISS
Eye-piece or ocular
Revolving nose
with objectives
Eye-piece or ocular
Revolving nose
with objectives
Stage and its controls
Focussing knob
Eye-piece or ocular
Revolving nose
with objectives
Stage and its controls
Condenser
Light source
Focussing knob
Light and lenses are the two pieces of equipment
which are used to manipulate the light. Both are
inherent to the microscope you used; they cannot
be changed.
The lenses: three types in your microscope
• The condenser
• The objective
• The eye-piece or ocular
The lenses: three types in your microscope
• The condenser
• The objective
• The eye-piece or ocular
The condenser: a combination of lenses
http://micro.magnet.fsu.edu/primer/anatomy/condensers.html
The condenser: a combination of lenses
http://www.zeiss.com/C1256B5E0047FF3F?Open
The lenses: three types in your microscope
• The condenser
• The objective
• The eye-piece or ocular
The objective: a combination of lenses
http://micro.magnet.fsu.edu/primer/
The objective: a combination of lenses
Motic Zeiss
http://micro.magnet.fsu.edu/primer/
The objective: a combination of lenses
Motic Zeiss
Plan CP-Achromat
http://micro.magnet.fsu.edu/primer/
Achromat:
Good color correction – exactly for two wavelengths.
Field flatness in the image center, refocusing also covers
the peripheral areas. Designed for fields of view up to
18 mm diameter.
Versions for phase contrast. Budget-priced objectives.
Names: CP-Achromat (CP: Clinical Plan) and
Achrostigmatism.
http://www.zeiss.com/C1256B5E0047FF3F?Open
The effect of chromatic aberration
Achromat: Zeiss
Good color correction – exactly for two wavelengths.
Field flatness in the image center, refocusing also covers
the peripheral areas. Designed for fields of view up to
18 mm diameter.
Versions for phase contrast. Budget-priced objectives.
Names: CP-Achromat (CP: Clinical Plan) and
Achrostigmatism.
http://www.zeiss.com/C1256B5E0047FF3F?Open
Field curvature: the sharpest focus of a lens is
on a curved surface rather than on a flat plane.
Motic Zeiss
Plan CP-Achromat
4x / 0.10
10x / 0.25
40x / 0.65
http://micro.magnet.fsu.edu/primer/
The objective: a combination of lenses
Motic Zeiss
Plan CP-Achromat
4x / 0.10 5x / 0.12
10x / 0.25 10x / 0.25
40x / 0.65 40x / 0.65
100x / 1.25 oil
http://micro.magnet.fsu.edu/primer/
The objective: a combination of lenses
Motic Zeiss
Plan CP-Achromat
4x / 0.10 5x / 0.12
10x / 0.25 10x / 0.25
40x / 0.65 40x / 0.65
100x / 1.25 oil
http://micro.magnet.fsu.edu/primer/
The objective: a combination of lenses
Motic Zeiss
Plan CP-Achromat
4x / 0.10 5x / 0.12
10x / 0.25 10x / 0.25
40x / 0.65 40x / 0.65
100x / 1.25 oil
∞ / 0.17 ∞/-
∞ / 0.17
http://micro.magnet.fsu.edu/primer/
All the objectives mentioned here are members of the family
of ICS-Optics (ICS: Infinity Color-corrected System). These
objectives project their images to “infinity” first. Only the tube
lens produces an intermediate image – to be more precise, at
a distance of approx. 164.5 mm behind the tube lens. This
distance was chosen to comply with the classical tube length.
The objective: a combination of lenses
Motic Zeiss
Plan CP-Achromat
4x / 0.10 5x / 0.12
10x / 0.25 10x / 0.25
40x / 0.65 40x / 0.65
100x / 1.25 oil
∞ / 0.17 ∞/-
∞ / 0.17
http://micro.magnet.fsu.edu/primer/
A coverslip is
Motic Zeiss
Plan CP-Achromat
4x / 0.10 5x / 0.12
10x / 0.25 10x / 0.25
40x / 0.65 40x / 0.65
100x / 1.25 oil
∞ / 0.17 ∞/-
∞ / 0.17
Any questions?
http://micro.magnet.fsu.edu/primer/
The lenses: three types in your microscope
• The condenser
• The objective
• The eye-piece or ocular
http://micro.magnet.fsu.edu/primer/anatomy/oculars.html
Zeiss
Designed for eyeglass wearers.
The exit pupil is at a considerable
PL 10x / 18 distance from the eyepiece.
Glasses symbol
Final image
Intermediate
image
Specimen
What is the main purpose of the microscope?
• The condenser
• The objective
• The eye-piece or ocular
What is the main purpose of the microscope?
• The condenser
• The objective
• The eye-piece or ocular
Intermediate
image
Specimen
Any questions?
What is the main purpose of the microscope?
http://micro.magnet.fsu.edu/primer/lightandcolor
Resolution: the minimum distance
between 2 dots that can be discerned
http://micro.magnet.fsu.edu/primer/
The smaller the diameter of the Airy disc produced by
a lens, the higher is the resolving power of that lens,
the better you can separate two distinct points.
http://micro.magnet.fsu.edu/primer/
Resolution = (0.61 λ) / Numerical Aperture
The larger the numerical aperture of a lens,
The smaller the Airy disc,
The smaller and better the resolution.
http://micro.magnet.fsu.edu/primer/
Airy disk sizes vary with changes in objective
numerical aperture and illumination wavelength.
λ: wavelength of light
n: refractive index of the medium in the object space
Θ: angular aperture
Resolution = (0.61 λ) / Numerical Aperture
Resolution = (0.61 λ) / (n x sin θ)
λ: wavelength of light
n: refractive index of the medium in the object space
Θ: angular aperture.
Objective
=θ
Specimen
where 0.61 represents the degree to which image points can overlap
and still be recognized by an observer as separate points
There is no diffraction
of light because of the
homogeneity of the
refractive indices.
http://micro.magnet.fsu.edu/primer/
Resolution = (0.61 λ) / (n x sin θ)
Any questions?
The objective: a combination of lenses
Motic Zeiss
Plan CP-Achromat
4x / 0.10 5x / 0.12
10x / 0.25 10x / 0.25
40x / 0.65 40x / 0.65
100x / 1.25 oil
∞ / 0.17 ∞/-
∞ / 0.17
r = (0.61 λ) / NA
http://micro.magnet.fsu.edu/primer/
It would be a pity if the intermediate
image produced with such sophisticated
optics were to be impaired just before it
reaches the eye because of poor handling
of the microscope.
Koehler illumination
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
• Place slide on stage
• Reduce brightness of light if need be
• Position the oculars so that you are
comfortable when looking through them
• Adjust your view by focussing the
diopter compensation ring
http://www.zeiss.com
Relaxed viewing is important
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
http://www.zeiss.com
Koehler or Köhler illumination
Maximum resolution
Maximum contrast
Maximum depth of detail
Creation of artefacts
Klosevych, 1989
Microscopy and photomicrography. R.F. Smith, CRC Press
1 3
http://micro.magnet.fsu.edu/primer
http://www.zeiss.com
(Microscopy from the beginning)