Introduction to Microscopy

1
Introduction to Microscopy
VIBS 443/602
2
Use your atlas!
Use your atlas!
Use your atlas!
Use your atlas!
pancreas
blood
stomach
Atlas of Normal Histology by DiFiore
testis
3
Getting Started in the Laboratory

• Light microscope parts
• page 24
• Kohler illumination pages 34-35

4
Kohler illumination (order is important)

1. Focus specimen
2. Close field diaphragm
3. Focus condenser
4. Center condenser
5. Open field diaphragm

5
Kohler illumination

1. Focus specimen
2. Close field diaphragm
3. Focus condenser
4. Center condenser
5. Open field diaphragm

6
MAGNIFICATION VS. RESOLUTION

• MAGNIFICATION - INCREASE IN IMAGE SIZE
• 2. RESOLUTION - SMALLEST DISTANCE BETWEEN TWO
  POINTS THAT CAN BE SEEN (DISTINGUISHED)
• CALCULATED BY
• 0.61 (WAVELENGTH)/NUMERICAL APERTURE
• 0.25 um FOR LIGHT MICROSCOPE
• 0.1 nm FOR ELECTRON MICROSCOPE

7
(No Transcript)
8
(No Transcript)
9
Peripheral blood smear (May-Grunwald-Giemsa)
113
10
Peripheral blood smear (Leishman-Giemsa)
110
11
Peripheral blood smear (Leishman-Giemsa)
110
12
Peripheral blood smear (Leishman-Giemsa)
110
13
Peripheral blood smear (Leishman-Giemsa)eosinoph
il, basophil, and neutrophils
110
14
Peripheral blood smear (Leishman-Giemsa)eosinoph
il, basophil, and neutrophils
110
15
(No Transcript)
16
Pancreas
158
17
Pancreas
158
18
Pancreas, monkey (toluidine blue)
156
19
Pancreas, monkey (toluidine blue)
156
20
(No Transcript)
21
UT165 larger secondary spermatocytes and smaller
Golgi phase spermatids human testis
165
22
Testis - blood and lymph vessels
19680
23
19709 Transparency of unstained tissue
24
19709 Transparency of unstained tissue
25
Mast cell in unstained tissue
19709
26
Mast cells in stained testicular tissue
19709
27
Mast cells in stained testicular tissue
19709
28
(No Transcript)
29
Fundic stomach
145
30
Fundic stomach
145
31
Argentaffin cells of Fundic stomach, rabbit
(toluidine blue)
244
32
Mucus neck cells Fundic stomach, rabbit
(toluidine blue)
244
33
EM 8f
EM 12a
EM 4c
EM 6a
Compare sizes of membranes ribosomes
mitochondria
EM 2b
EM 7
34

• EM 8f Peripheral blood cells 9,000x
• Monocyte
• Lymphocyte
• Neutrophil

35

• EM 12a Bone marrow 13,200x. Note the reticular
  cell and developing red blood cells.
• Reticular cell
• Developing red blood cell

36

• EM 4c Intestinal absorption cell 60,000x
• Budding RER
• Coated vesicle
• Golgi
• Mitochondria
• Nucleus
• Plasma membrane
• Primary lysosome

37

• EM 2b Liver 60,000x cytoskeletal elements.
  Microtubes,
• microfilaments, and intermediate filaments can
  be compared in this
• cell, which has a high concentration of cortical
  microfilaments.
• Microtube
• Microfilaments
• Intermediate filaments

38

• EM 7 Ascites fluid 80,000x. Clear examples of
  Golgi apparatuses with their cisternae and
  vesicles are present in this cell
• Golgi apparatus
• Ribosomes
• Lipofuscin
• Mitochondrion

39

• EM 6a Centriole-microtubules 200,000x.
  Centriolar region of a cell showing both the
  stable, triplet microtubule arrays within the
  centriole, and the labile, individual
  microtubules originating from pericentriolar
  material.
• Centriole
• Stable microtubule
• Labile microtubule

40
EM 4c 60,000x
EM 8f 9,000x
Compare sizes of membranes ribosomes
mitochondria
EM 12a 13,200x
41
EM 6a 200,000x
EM 2b 60,000x
Compare sizes of membranes ribosomes
mitochondria
EM 7 80,000x
42
Conventional TEM, SEM carbon
replica TEM
EM 2b
EM 4a
EM 18b
EM 6
EM 7
EM 8
43
(No Transcript)
44
(No Transcript)
45
(No Transcript)
46
(No Transcript)
47
Conventional TEM, SEM carbon
replica TEM
EM 2b
EM 4a
EM 18b
EM 6
EM 7
EM 8
48
Conventional TEM, SEM carbon
replica TEM
EM 2b
EM 4a
EM 18b
EM 6
EM 7
EM 8
49
Conventional TEM, SEM carbon replica
TEM
EM 2b
EM 4a
EM 18b
EM 6
EM 7
EM 8
50
Use your atlas!
Use your atlas!
pancreas
blood
stomach
Atlas of Normal Histology by DiFiore
testis