LOGO MLT 1222 Microbiology Instrumentation By: Norazli Ghadin

Lesson Objectives 1. Introduce to basic microbiology instrument 2. Know how to use instrument 3.Naming parts of microscope 4.Know the function of microscope parts 5.Know how to observe microbiology slides using microscope

The instrument  In microbiology the instruments are designed to  Maintain sterility  To provide easy manipulation of bacterial culture  To maintain pure culture  To ensure bacterial growth are controlable

Biosafety cabinet Enclosed, ventilated workspace for safely working with materials contaminated with (or potentially contaminated with) pathogens There are 3 level of biosafety cabinet 1. Biosafety level 1 2.Biosafety level 2 3.Biosafety level 3

Biosafety cabinet  Works Principle  Provide protection to operator and sample(class II & III)  Air came out of cabinet are filtered,  Remove microbe

Biosafety cabinet  Biosafety level I  Protect operator  Air entering did not HEPA filtered  Air coming Out are filtered. Samples e.g  Special equipment  Culture aeration

Biosafety cabinet  Biosafety level II  Protect operator and samples  Incoming air and waste air is HEPA filtered  Samples All types of microorganisms exclude virus, volatile chemicals, gases

Biosafety cabinet  Biosafety level III  Protect operator and samples  The enclosure is gas-tight.  All materials enter and leave through a dunk tank or double-door autoclave.autoclave  Gloves attached to the front prevent direct contact with hazardous materials.  the lab equipment installed inside custom-built as well.

Biosafety cabinet

 Samples  Highly contagious  Such as Ebola, air borne virus, H1N1

Microbiology Incinerator  Use for sterilizing wire loop  Electrical change to heat  High temp destroys living organisms

Bunsen burner  Alternative for microbiological incinerator

Wire Loop/Stab  For inoculating microbes  Sterilize with microbe incinerator

PETRI DISH  Use to culture microbe  Fill with media such as nutrient agar

Bijou bottle  Use to culture microbe on Slant agar/liquid broth

M ICROSCOPE SLIDE  Use to put on sample  Prepare microbe for staining and microscope observation

Conical FLASK  For culturing microbes  Cotton gauze,  Membrane filtered cap.

Staining Rack  To put glass slide for staining.

 To prepare media, solution, or broth in large volume.  High resistant to temperature and preasure

Culture Tube  For culturing microbes  Small volume solution.  Can also use 96 wells  plate

 A microscope (from the Greek: μικρός, mikrós, "small" and σκοπε ῖ ν, skopeîn, "to look" or "see") is an instrument to see objects too tiny for the naked eye.Greekinstrument  The science of investigating small objects using such an instrument is called microscopy. Microscopic means invisible to the eye unless aided by a microscope.microscopy Microscopic LAB INSTRUMENTATION MLT 1102 Microscope

 Eyepiece:  Magnifies material being viewed by 10X  The part of the microscope you look into  Sometimes contains a pointer that can be seen as you look into the eyepiece.  May also be called the ocular. LAB INSTRUMENTATION MLT 1102 Component

 Nose piece:  Part of microscope to which the objectives are attached  Rotates to allow for the changing of objectives to increase or decrease magnification. LAB INSTRUMENTATION MLT 1102

 Arm:  Part of microscope to which the nosepiece is attached  A secure part of the microscope to hold on to when the microscope is being carried.  Sometimes includes a cord hanger for power cord storage LAB INSTRUMENTATION MLT 1102

 Objectives:  Magnify material being examined  Scanning magnifies 4X  10X magnifies 10X  High dry magnifies 40X  Oil immersion magnifies 100X Total magnification = magnifying power of eyepiece times the magnifying power of the objective in use. LAB INSTRUMENTATION MLT 1102

 Stage:  Platform on which microscope slide rests Mechanical Stage: Used for adjusting the position of the slide for viewing Used for adjusting the position of the slide for viewing Slide holder: Secures slide to the mechanical stage Secures slide to the mechanical stage Mechanical Stage control knobs: Move the slide Move the slide LAB INSTRUMENTATION MLT 1102

 Coarse adjustment knob:  Coarse focusing Fine adjustment knob: Precise focusing Precise focusing LAB INSTRUMENTATION MLT 1102

 Substage condenser  concentrates light & directs it through opening in microscope stage Iris diaphragm: regulates the amount of light passing through the substage condenser regulates the amount of light passing through the substage condenser increases resolving power of the microscope increases resolving power of the microscope LAB INSTRUMENTATION MLT 1102

Iris diaphragm lever: adjusts the iris diaphragm to increase or decrease microscope resolution adjusts the iris diaphragm to increase or decrease microscope resolution Condenser control knob: raises & lowers condenser raises & lowers condenser LAB INSTRUMENTATION MLT 1102

 Illuminator  Light source Illuminator control: Controls the intensity of light Controls the intensity of light Serves as the on/off switch for some microscopes Serves as the on/off switch for some microscopes Base: provides support for microscope provides support for microscope LAB INSTRUMENTATION MLT 1102

Using Microscope  Put stained slide on stage, secure with clip  Rotate nosepiece so 4X objective face the slide,  Use coarse focusing to focus the image,  Change to 10X objective, use fine focusing to get sharp image.  Change to 40 X use fine focus knob  Rotate objective to 100X halfway,  Drop an immersion oil to the slide  Continue rotating the nosepiece to 100X objectives.  Slowly use fine focusing to get the sharp image.