LAB SAFETY

universal precaution

All cultures shouldbe handled with respect and proper aseptic technique as if they were potential pathogens. This is called

LAI - LABORATORY ACQUIRED INFECTION

infection that prevent in lab

NOSOCOMIAL

HOSPITAL INFECTION

STERILE

IT HAS NO MICROORGANISMS

AUTOCLAVING

STEAMED UNDER PRESSURE

ANTON VAN LEEUWENHOEK

FATHER OF PROTOZOLOGY

GLASSWARE

TOOLS

EQUIPMENT

A MICROBIOLOGY LABORATORY REQUIRES WELL-BUILT ROOM WITH ________, _______, _____

testtubes

culture tubes

petri dishes

graduated cylinders

pipettes

flasks

The most important glassware used in a microbiological laboratory are _______, __________, _________, ___________, _______, _______

inoculation needle

inoculation/transfer loop

Bunsen burner

autoclave (or pressure cooker)

incubator

hot air oven

refrigerator

centrifuge

spectrophotometer

hot plate

UV- lamp

magnetic stirrer

water-bath

single-pan balance with weights (forrough use)

pH meter

laminar air flow

microscopes

The most common instruments are an

MICROSCOPE

• used to view microscopic organisms or life forms that cannot be seen with the naked eye

• Used primarily for the initial direct identification of bacteria based on Gram stain characteristic, as well as their morphological features

shape

size

arrangement

staining reaction

microscope used in initial identification of bacteria based on:

gram staining

staining reaction common in bacteria is

need siya istain kasi pag walang kulay mababa contrast sa microscope and hindi siya makikita

why need istain

Bright-Field/Light Microscope

Phase-contrast Microscope

Fluorescence Microscope

Electron Microscope

TYPES OF MICROSCOPES

Bright-Field/Light Microscope

- commonly used in clinical and teaching laboratories that usually magnifies an object up 1000x (diameters) (STAINED SPECIMEN)

1000x or minsan 2000x

diameters of bright field/light microscope

Phase-contrast Microscope

- permits one to see internal details such as the nucleus, cytoplasm and granules of unicellular organisms (UNSTAINED SPECIMEN)

parasites and urine para makita yung activities

diff direction

phase contrast microscope used in ________,_____ and the light if direct in ________

Fluorescence Microscope

- utilizes ultraviolet light, fluorescent dyes and on occasion antibody molecules. The microbial specimen being observed appears to glow in the dark (SPECIAL STAINING)

FLUORESCENT DYES

UV LIGHT

SPECIAL STAINING TECHNIQUE THAT USED __________ (PANG EXCITE =________)

fluorenscence microscope

treponema

it used para maging specific yung bacteria like

Electron Microscope

COATING

metal for organism

- uses a beam of electrons as its source of energy that detects very tiny objects such as viruses or molecules

AND IT USE ________ with positive charge particle like ____________

electron microscope

it use in nobel organism or unknown

can see molecules (structure of molecule)

doesnt use stains

dapat malaman to para magroup

SCANNING/TUNNELING MICROSCOPE

TRANSMISSION MICROSCOPE

2 ELECTRON MICROSCOPE

SCANNING/TUNNELING MICROSCOPE

OUTER LANG NG BACTERIA MAKIKITA

TRANSMISSION MICROSCOPE

INSIDE THE BACTERIA

BROTH

CALLED IN LIQUID TYPE OF CULTURE

INOCULATING LOOP/WIRE

are hand-held and compact appliances that introduce microorganisms like bacteria or yeast into plated or tubed growth media before incubation, multiplication, or growth

INOCULATING LOOP/WIRE

TRANSFERRING DEVICE CAN BE MADE UP OF METAL (REUSABLE) OR PLASTIC (DISPOSABLE) TRANSFER SOLID OR LIQUID TYPE OF CULTURE

LOOP

WIRE

2 TYPE OF INOCULATING

Loops

Wires

- used to retrieve liquid media

- used to retrieve solid or dense media

LOOP OR NEEDLE

WHAT INOCULATING USED IN SOLID CULTURE

LOOP

WHAT INOCULATING USED IN LIQUID CULTURE

1. Calibrated loop

2. Non-calibrated loop

3. Reusable (Wire) needle

4. Disposable needle

Types of Inoculating Loops/Needles

LOOP AND WIRE

PART OF INOCULATING LOOP/WIRE

CALIBRATED LOOP

HAVE SPECIFIC VOLUME

1 UL & 10 UL

COMMON VOLUME OF CALIBRATED LOOP

QUANTITATIVE CULTURE

CALIBRATED LOOP

IT COUNT THE CULTURE AND IT USED ________

NON-CALIBRATED

IT HAS NO SPECIFIC VOLUME

BUNSEN BURNER/ALCOHOL LAMP

use for sterilizing inoculating loop, plating out cultures, heat-fixing of smears and creating zone for aseptic operation

HOTPLATES

is used to heat glassware or its contents specially when preparing culture media which requires homogenization

providr distributed type of heat

PETRI DISH

are made of plastic or glass used to hold media and grow bacteria for study.

• Use to hold small objects for observation

• Used in the evaporation and drying of samples

50ml (common)

150ml

size of petri dish

small object

solid culture

petri dish hold

plated

tube

type of media

solid

plated

solid

liquid (broth)

semi solid

tube

part of petri dish

inverted position (top down)

TAKE NOTE:⚬ Petri dishes are incubated in an __________ to preventcondensation formed on the cover during solidification from dropping down ontothe surface of the hardened agar.

TEST TUBES

used to cultivatemicroorganisms.

broth or agar

TEST TUBES A suitable nutrient medium in the form of ________ may be added to the tubes.

sterile environment

A _____________ is maintained in culturetubes by various types of closures.

Schröeder and von Dusch

nineteenth century

Historically, the first type, a cotton plug, was developed by ________,__________ inthe ____________.

bacteriological tube

test tube aka

solid

semi solid

liquid

test tube used to hold the culture like

bacteriological tube

screw cap

plastic closure

metal closure

nonabsorbent cotton

what name

Morton closures

Today most laboratories use sleevelike caps made of metal, such asstainless steel, or heatresistant plastics.

morton closures

The advantage of these closures over thecotton plug is that they are laborsaving and,most of all, slip on and off the test tubeseasily.

ERLENMEYER FLASKS

are used to contain liquids and for mixing, heating, cooling, incubation, filtration,storage, and other liquid-handling processes.

swirling without the risk of spills

titrations and for boiling liquids.

Their slanted sides and narrow necks allow thecontents to be mixed by ______________ , which is useful for _____________

FORCEPS

sterile forceps

In fields like microbiology or clinical laboratories, _________ are used to handlesterile materials, such as agar plates, sterile swabs, or sterile packaging, to maintain aseptic conditions.

forceps

antibiotic disc

manipulates small objects like

ANALYTICAL BALANCE

• use for the accurate weighing of samples, media, chemicals and precipitates

• use to measure mass with high degree of precision

AUTOCLAVE

devise use to sterilize equipment and supplies by subjecting them to specified temperature, pressure and time

• it is the pre-disposal treatment and sterilization of waste material

121°c

15psi/1 qtm

15-30mins (common in glassware)

common temperature to kill bacteria and pressure, time

35-37°c

temp to grow mostly the mdecially bacteria

geobacillus stearothermophillus spores

what bacteria hindi agad namamatay sa 121c temp

yellow color change

failure

if the Geobacillus stearothermophilus detected by a visual color change (media turn yellow) the ___________ indicates a sterilization process _______

incubator (tamang temp and gas requirement)

devise use to grow and maintain microbiological culture or cell culture and maintains optimal temperature and other conditions such as CO2 and O2 content of the atmosphere

BIOSAFETY CABINET (BSC)

• enclosed, ventilated laboratory workplace for safety working with materials contaminated with pathogens requiring a defined biosafety level

• use to protect the laboratory worker and the surroundings

class I

class II (class IIA1, class IIA2, class IIB1, class IIB2)

class III

3 type of biosafety cabinet

CLASS II

USUALLY MAKIKITA SA HOSPITAL

CLASS IIA2

COMMON TYPE OF CLASS II

REF

ANTIBIOTIC

SERUM

BIOCHEMICAL REAGENT

MAINTAIN OR STORED NG CULTURE/STOCK CULTURE FOR _________,_______,_______

REFRIGERATOR

is used for a wide variety ofpurposes such as maintenance andstorage of stock cultures betweensubculturing periods, and storage ofsterile media to prevent dehydration. It isalso used as a repository for thermolabilesolutions, antibiotics, serums, andbiochemical reagents

STOCK CULTURE

SEPARATE PART OF MAIN CULTURE

COLONY COUNTER

used to count colonies of bacteria or other microorganisms growing on an agar plate

QUANTITATIVE CULTURE

COLONY COUNTER IS FOR

URINE CULTURE

COLONIE FORMING UNITS PER ML FOR_______

• Concentrated bleach in a working dilutionof 1:10 dilution of bleach

• 70% Alcohol

• Disinfectant bottle container with clean water

• Tissue

• Biohazard bag

• PPEs

• Alcohol-based sanitizer

BIOLOGICAL SPILL RESPONSE

REPORT

HAND HYGINE

DON PPE

PERFORM THE ACTUAL SPILL PROCEDURE

STEP IN SPILL

1:10 dilution of bleach

Concentrated bleach in a working dilution of __________