BacteLab Preliminary PDF

MICROSCROPE 12. Stage clips – hold slide in place - Op#cal instrument that uses lens or combina#on of 13. Aperture – hole in stage through which base light lenses to produce magnified images of small objects,...

MICROSCROPE 12. Stage clips – hold slide in place - Op#cal instrument that uses lens or combina#on of 13. Aperture – hole in stage through which base light lenses to produce magnified images of small objects, reaches the stage especially of objects to be seen by the unaided eye 14. Condenser – collect and focus light on stage. - Used to see bacteria whose: Located under the stage ohen in conjunc#on with average size is: 0.5 to 2.0 μm an iris diaphragm average diameter range from 0.3 to 1.0 μm Types of microscope Resolu#on 1. Simple microscope – early microscope - Known as “Resolving power” 2. Compound microscope – has 2 lenses and - Ability of lenses: to reveal detail or 2 points illumina#on system (sun &mirror/lamp) dis#nctly separated 3. Monocular – 1 eyepiece - Func#on of wavelength of light used 4. Binocular – 2 eyepiece - Characteris#cs of lens system called numerical Basic frame of the Microscope aperture 1. Stage - Resolving power = wavelength of light used 2. Tube 2 x numerical aperture 3. Arm 4. Base Spherical Aberra#on - When you bring the center of the microscope field Microscope parts and func#ons: into focus, the peripheral may be fuzzy due to 1. Eyepiece – where you look to see image of your curvature of the lens, resul#ng in mul#ple focal specimen points 2. Base – boTom of microscope, used for support - This can be minimized by the use of iris diaphragm 3. Arm – supports tube and connects it to the base 4. Objec#ve lenses – increase magnifica#on of Chroma#c Aberra#on specimen - Mul#tudes of colors a. Scanning objec#ve lens - Prism like effect of the lens as various wavelength - Magnifies 4x of white light pass through to a different focal - Shortest objec#ve point for each wavelength - Used for general overview of slide - Can be minimized by the use of filters (Blue) b. LPO/Low power objec#ve – magnify 10x a. Achroma#c lens c. HPO/High power objec#ve – magnify 40x - Lens corrected for red & blue d. OIO/Oil immersion objec#ve (white) light - Magnify 100x b. Apochroma#c aberra#on - Used wet either: - Lens correct for red, blue, and Cedar wood oil other wavelength Synthe#c oil c. Monochroma#c light - Most logical but most expensive Objec#ves Magnifica#on Eyepiece mag. Total mag. method of elimina#ng chroma#c Scanner 4x 10x 40x aberra#on LPO 10x 10x 100x HPO 40-44x 10x 400-440x EXAMINATION OF LIVING ORGANISMS OIO 97-100x 10x 970-1000x - Anton van Leewenhoek TOTAL MAGNIFICATION = Objec#ve lens x Eyepiece o First known individual to observe living microbes in a suspension Example: - Wet mount techniques are used for studying What is the total magnifica#on of LPO? microbes in its living state Total magnifica#on = 10(LPO) x 10(eyepiece) - Most bacteria die rapidly if no moisture is available = 100x for them - Microbes will exhibit either 5. Iris diaphragm – control light going through o Brownian movement aperture o True mo#lity 6. Light source – reflect light to specimen 7. Body tube – connects eyepiece to objec#ve lenses 2 types of prepara#on used to examine living organisms 8. Revolving nose piece – holds two or more 1) Wet mount objec#ve lenses and rotated to easily change 2) Hanging drop prepara#on power 9. Stage – flat plaform where you place your slides - Direct examina#on of living organism is very useful in 10. Fine adjustment knob – small knob used to fine- determining: tune the focus of your specimen a) Size 11. Coarse adjustment knob – Large knob used for b) Shaped focusing specimen c) Movement  Acidic stain – acid fuchsin, eosin Hay Infusion - Excellent way to provide microbes at a high density during any #me of the year DIFFERENT KINDS OF STAIN - Allows to follow development of microbes as they 1. Simple stain – one dye or reagent become visible over #me a. Direct – stains bacteria b. Indirect / nega