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CHAPTER 3 OBSERVING MICROORGANISMS THROUGH A MICROSCOPE

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Presentation on theme: "CHAPTER 3 OBSERVING MICROORGANISMS THROUGH A MICROSCOPE"— Presentation transcript:

1 CHAPTER 3 OBSERVING MICROORGANISMS THROUGH A MICROSCOPE
Dr. Reitano SUFFOLK COUNTY COMMUNITY COLLEGE

2 Microorganisms were first observed by Antonie van ____________, using a ________ microscope. A _____________microscope has only one lens. Cowan “Microbiology”

3 RELATIVE SIZE as a TOOL in ________________
_____ System – International System of Units Cowan “Microbiology”

4 SOME TYPES of MICROSCOPES
________ Bright field __________ LIGHT MICROSCOPE _____________ ELECTRON Transmission Scanning

5 ________________ MICROSCOPY
DISTINGUISHING FEATURES Visible light passes through a series of ________ which magnify the specimen and allows fine detail to be observed (___________) Specimen appears _______ Field appears lighter PRINCIPLE USES Common, multi-purpose microscope Used to observe ____ specimens and preserved, stained (___-living) specimens Provides fair cellular detail

6 _________________ MICROSCOPE

7 ___________ Light Microscopy
PATHWAY of _____ In a compound microscope, the image from the ____________ lens is magnified again by the __________ lens Total magnification = objective lens  ocular lens Figure 3.1b

8 Properties of a Compound Light Microscope: 1. _____________________
Magnification of _________ Lens X Magnification of Ocular Lens = Total Magnification Low Power ___X 10X 100X High Dry 400X Oil Immersion 1000X 2. _____________ The ability of a lens system to accurately distinguish between two separate points, that lie close to each other, as separate and distinct. (structures less than 0.2um cannot be resolved with the compound light microscope)

9 _________ in the Compound Microscope
______ index - a measure of the light-bending ability of a medium, such as air Air may bend the light so much that it misses the small size of the opening in the 100x objective lens ____________ is used to keep light from bending Figure 3.3

10 _______________ MICROSCOPY
DISTINGUISHING FEATURES Specimens are stained with ____________dyes ultraviolet light is used which causes ___________ molecules in a specimen to emit light PRINCIPLE USES Rapid detection and identification of organisms in tissues Excellent _____________ tool

11 ELECTRON MICROSCOPE TYPES: _____________________ ___________________________

12 __________ ELECTRON MICROSCOPY
Principle Uses Observing _______ details of cells and viruses DISTINGUISHING FEATURES Beam of __________(not light) are reflected from the specimen __________dimensional image produced Magnification 1,000 to 10,000x

13 ___________ ELECTRON MICROSCOPY
DISTINGUISHING FEATURES Beam of __________ (not light) pass through the specimen ___ dimensional image is produced Magnification 10,000 to 100,000x PRINCIPLE USES Examination of _______ _____________ of cells

14 COMPARISON of _____ MICROSCOPES and ____________ MICROSCOPES
Cowan and Talaro

15 MICROSCOPIC OBSERVATION
TWO TYPES of SPECIMENS: 1. ____________ Wet Preps, Wet Mounts. Living organisms suspended in fluid-organisms have ________________with surrounding fluid. Used to study: size, shape, arrangement of cells, (morphology), __________ , and _________. 2. STAINED Fixed Smear Preparations. ________________ organisms. Contrast is created to allow cellular characteristics to stand out. Used to study size, shape, arrangement of cells (morphology) but not motility.

16 PREPARATION of _______ SMEAR
SMEAR: A ___ film of a solution of ________ on a slide A smear is usually ____ to ______ the microbes to the slide and to ______ the microbes. ______: Coloring the microbe with a ____ that emphasizes certain structures _________ STAINING Nester et al.

17 VARIOUS STAINING CATEGORIES
ACIDIC STAINS __________ charge (negative stain) _________ by cells _________ is stained Ex.: India ink ________ stain BASIC STAINS ________ charge (positive stain) ________ to cells (cells have a _______ charge) Cells are stained Ex.: Methylene Blue, Crystal Violet, Safranin, Malachite Green

18 VARIOUS STAINING CATEGORIES cntd.
SPECIAL Targets _______ cell _______ Such as: capsules flagella spores Ex.: India Ink Flagella Stain Spore Stain SIMPLE _____ dye Simple procedure Ex.: Methylene Blue Crystal Violet Safranin Malachite Green DIFFERENTIAL Two dyes __________ Contrast 2 cell types or parts Complex procedure Ex.: _____ Stain ___________ Stain

19 _______________ STAIN GRAM STAIN
Developed by Dr. Hans Christian Gram in 1884 Most widely used procedure for staining ___________ Classifies bacteria into two groups Based on differences in _________ STRUCTURE Gram positive Gram negative

20 Basic _____ of Most Bacterial Cell _____
Basic _______ of Bacterial Cell Walls: determine _______ provide structural _________

21 COMPARISON of GRAM POSITIVE and GRAM NEGATIVE BACTERIA
Ex.: Staphylococcus aureus Streptococcus pyogenes GRAM _________ BACTERIA Ex.: Escherichia coli Klebsiella pneumoniae Nester

22 Gram-positive bacterial cells Gram-negative bacterial cells
Gram Stain PROCEDURE STEPS: Color of Gram-positive bacterial cells Gram-negative bacterial cells 1. ________ stain: Crystal Violet Purple 2. ____________: Iodine 3. _____________ agent:** Acetone Alcohol _________ 4. Counterstain: _____________ ___________ ________

23 _______ STAINING PROCEDURE
Tortora

24 ________________ STAIN Acid Fast Stain
Used for bacteria with _______ material in cell wall Several procedures* Ex.: Mycobacteria species Nocardia species _____bacteria species Nester

25 DIFFERENTIAL STAIN ________________
Used for bacteria with waxy, lipid (_____ ______) material in cell wall Several procedures* 1. Primary stain: Carbolfuchsin 2. Decolorizer: _____ Alcohol 3. Counter stain: Methylene blue Ex.: Mycobacteria species Nocardia species Mycobacteria species Nester

26 ___________ Stain Procedure
STEPS: Color of ___________ Bacteria (Mycobacteria sp.) Non–Acid-fast Bacteria 1. Primary stain: __________ Red 2.__________ agent:** _____-alcohol ___________ 3. Counter-stain: _____________ ____ ______

27 ACID FAST STAIN Acid-fast staining of a patient’s ________ is a rapid, reliable, and inexpensive method to diagnose __________. What is the genus and species of this organism? This is an acid-fast stain of a patient’s _______. What is the disease associated with this organism?

28 SPECIAL STAINS Used to distinguish _____ of cells
CAPSULE ______________ ENDOSPORE Tortora

29 COMPARISON of STAINS Cowan et al.

30 SOME IMPORTANT STAINS USED in MICROSCOPY
________ - Methylene Blue - Carbolfuchsin - Crystal Violet - Safranin DIFFERENTIAL - Gram - Acid-Fast SPECIAL - __________ (Negative, Acidic) - Endospore - Flagella


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