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Dr. Sudha Kumari
Assistant Professor
Department of Veterinary Microbiology
Bihar Animal Sciences University, Patna
MICROSCOPY AND MICROMETRY
INTRODUCTION
DEFINITION HISTORY
BACKGROUND
VARIBLES USED
IN
MICROSCOPY
TYPES OF
MICROSCOPES
COMPOUND
MICROSCOPE -
Structure and
Function
USE OF
MICROSCOPE
CARE OF
MICROSCOPE
INTRODUCTION
Microscope is a precision instrument which provides
magnification so that it enables us to see an object and its structure
otherwise invisible to the naked eye.
Generally the microorganisms are smaller than this and thus a
microscope is essential for their observation.
DEFINITION
A microscope ( Greek : micron = small and scopes = aim)
MICROSCOPE - An instrument for viewing objects that are too small
to be seen by the naked or unaided eye.
MICROSCOPY - The science of investigating small objects using such
an instrument is called microscopy. OR Micrometry is the measurement
of microscopic objects (microorganisms).
HISTORICAL BACKGROUND
 1590 - Hans Janssen and his son Zacharias Janssen, developed first
microscope.
 1609 - Galileo Galilei - occhiolino or compound microscope.
 1620- Christian Huygens, another Dutchman, developed a
simple
2-lens ocular system that was chromatically corrected.
 Anton van Leeuwenhoek (1632-1723)
Anton van Leeuwenhoek is generally credited with bringing the
microscope to the attention of biologists.
A tradesman of Delft, Holland.
1661 - He discovered bacteria, free-living and parasitic
microscopic protests', sperm cells, blood cells, microscopic
nematodes etc.
 Microscope used by Anton von Leeuwenhoek An old pocket
Microscope.
VARIABLES USED IN Microscopy
MAGNIFICATION
Degree of enlargement
 No. of times the length, breadth or diameter, of an object is
multiplied.
RESOLUTION – Ability to reveal closely adjacent structural details as
separate and distinct.
LIMIT OF RESOLUTION (LR) – The min distance between two
visible bodies at which they can be seen as separate and not in contact
with each other.
TYPES OF MICROSCOPE:-
• Simple microscope
• Compound microscope
• Phase Contrast Microscope
• Dark Ground Microscope
• Fluorescent Microscope
• Electron Microscope
• Others
Parts of microscope
• Illuminator - This is the light source located below the specimen.
• Condenser - Focuses the ray of light through the specimen.
• Stage - The fixed stage is a horizontal platform that holds the
specimen.
• Objective - The lens that is directly above the stage.
• Nosepiece - The portion of the body that holds the objectives over the
stage.
• Iris diaphragm - Regulates the amount of light into the condenser.
• Base – Base supports the microscope which is horseshoe shaped.
• Coarse focusing knob - Used to make relatively wide focusing
adjustments to the microscope.
• Fine focusing knob - Used to make relatively small adjustments to
the microscope.
• Body - The microscope body.
• Ocular eyepiece - Lens on the top of the body tube. It has a
magnification of 10× normal vision.
OBJECTIVE LENS
Mounted on nose piece
It forms magnified real image.
Magnification of objective
= Optical Tube length
Focal Length
Types
1. Scan - 4X , 2. Low Power 10X
3. High Power - 40X , 4. Oil immersion - 100X
OIL IMMERSION OBJECTIVE
Highest magnification
Oil prevents refraction of light outwards and allows it to pass straight
in to objective.
USING COMPOUND MICROSCOPE
 Grasp the microscopes arm with one hand and place your other
hand under the base .
 Place the microscope on a bench.
 Adjust seat. Clean Lenses.
 Turn the coarse adjustment knob to raise the body tube arm.
 Revolve the nose piece to set low-power objective lens.
 Adjust the Condenser lenses and diaphragm.
 Place a slide on the stage and secure with stage clips.
 Switch on the light at low intensity and then increase intensity.
 Turn the coarse adjustment knob to lower the body tube until the
low power objective reaches its lowest point.
 Looking through the eyepiece, very slowly move the coarse
adjustment knob until the specimen comes into focus.
 Adjust distance between eye piece.
 Switch to the high power objective lens only after adjusting
condenser and iris diaphragm.
 Place a drop of oil over specimen before using oil immersion
objective.
 Lower the objective until oil makes contact with objective.
 Looking through the eyepiece, very slowly focus the objective
away from the slide.

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Microscopy-1.pptx

  • 1. Dr. Sudha Kumari Assistant Professor Department of Veterinary Microbiology Bihar Animal Sciences University, Patna MICROSCOPY AND MICROMETRY
  • 2. INTRODUCTION DEFINITION HISTORY BACKGROUND VARIBLES USED IN MICROSCOPY TYPES OF MICROSCOPES COMPOUND MICROSCOPE - Structure and Function USE OF MICROSCOPE CARE OF MICROSCOPE
  • 3. INTRODUCTION Microscope is a precision instrument which provides magnification so that it enables us to see an object and its structure otherwise invisible to the naked eye. Generally the microorganisms are smaller than this and thus a microscope is essential for their observation.
  • 4. DEFINITION A microscope ( Greek : micron = small and scopes = aim) MICROSCOPE - An instrument for viewing objects that are too small to be seen by the naked or unaided eye. MICROSCOPY - The science of investigating small objects using such an instrument is called microscopy. OR Micrometry is the measurement of microscopic objects (microorganisms). HISTORICAL BACKGROUND  1590 - Hans Janssen and his son Zacharias Janssen, developed first microscope.  1609 - Galileo Galilei - occhiolino or compound microscope.
  • 5.  1620- Christian Huygens, another Dutchman, developed a simple 2-lens ocular system that was chromatically corrected.  Anton van Leeuwenhoek (1632-1723) Anton van Leeuwenhoek is generally credited with bringing the microscope to the attention of biologists. A tradesman of Delft, Holland. 1661 - He discovered bacteria, free-living and parasitic microscopic protests', sperm cells, blood cells, microscopic nematodes etc.  Microscope used by Anton von Leeuwenhoek An old pocket Microscope.
  • 6. VARIABLES USED IN Microscopy MAGNIFICATION Degree of enlargement  No. of times the length, breadth or diameter, of an object is multiplied. RESOLUTION – Ability to reveal closely adjacent structural details as separate and distinct. LIMIT OF RESOLUTION (LR) – The min distance between two visible bodies at which they can be seen as separate and not in contact with each other.
  • 7. TYPES OF MICROSCOPE:- • Simple microscope • Compound microscope • Phase Contrast Microscope • Dark Ground Microscope • Fluorescent Microscope • Electron Microscope • Others
  • 8.
  • 9.
  • 10. Parts of microscope • Illuminator - This is the light source located below the specimen. • Condenser - Focuses the ray of light through the specimen. • Stage - The fixed stage is a horizontal platform that holds the specimen. • Objective - The lens that is directly above the stage. • Nosepiece - The portion of the body that holds the objectives over the stage. • Iris diaphragm - Regulates the amount of light into the condenser. • Base – Base supports the microscope which is horseshoe shaped.
  • 11. • Coarse focusing knob - Used to make relatively wide focusing adjustments to the microscope. • Fine focusing knob - Used to make relatively small adjustments to the microscope. • Body - The microscope body. • Ocular eyepiece - Lens on the top of the body tube. It has a magnification of 10× normal vision.
  • 12. OBJECTIVE LENS Mounted on nose piece It forms magnified real image. Magnification of objective = Optical Tube length Focal Length Types 1. Scan - 4X , 2. Low Power 10X 3. High Power - 40X , 4. Oil immersion - 100X
  • 13. OIL IMMERSION OBJECTIVE Highest magnification Oil prevents refraction of light outwards and allows it to pass straight in to objective. USING COMPOUND MICROSCOPE  Grasp the microscopes arm with one hand and place your other hand under the base .  Place the microscope on a bench.  Adjust seat. Clean Lenses.  Turn the coarse adjustment knob to raise the body tube arm.
  • 14.  Revolve the nose piece to set low-power objective lens.  Adjust the Condenser lenses and diaphragm.  Place a slide on the stage and secure with stage clips.  Switch on the light at low intensity and then increase intensity.  Turn the coarse adjustment knob to lower the body tube until the low power objective reaches its lowest point.  Looking through the eyepiece, very slowly move the coarse adjustment knob until the specimen comes into focus.  Adjust distance between eye piece.
  • 15.  Switch to the high power objective lens only after adjusting condenser and iris diaphragm.  Place a drop of oil over specimen before using oil immersion objective.  Lower the objective until oil makes contact with objective.  Looking through the eyepiece, very slowly focus the objective away from the slide.