INSTRUMENTS

DR. SALEHA MOMIN

ASSISTANT PROFESSOR
INOCULATING LOOP/
Nichrome wire loop
USES:
1. Inoculation of clinical specimens on solid or
liquid medium
2. To transfer the bacterial growth from one
medium to other medium to elicit
biochemical reaction
Sterilization :- making it red hot under Bunsen-
burner
Sterile Cotton Swab
Uses:
1. For collection of specimens from throat, cervix, local
leasions
2. For preparing lawn culture of bacterial growth as in
antibiotic sensitivity testing
Sterilization:- Hot air oven
Universal container
Uses :
1. For specimen collection such as urine, pus, body fluid atc.
2. For one time use only and then disposed of
Sterilization :-
i. By ethylene oxide
ii. By gamma radiation
Autoclave
Working Principle :- 121°C for 15 min at a pressure of 15 pounds (lbs)
Sterilization principle: moist heat sterilization method
Uses : for sterilization of culture media, rubber material, gloves,
gowns, dressing etc
Precautions
1. It should not be used for sterilizing waterproof materials such as
oil and grease
2. The chamber should not be overfilled and the material should
not touch the sides or top of the chamber
Sterilization Control
i. Biological indicator: Spores of Geobacillus stearothermophilus
ii. Chemical indicators : External pack control, e.g. autoclave tape,
Bowie-Dick test, Internal pack control
 Inspissator
Working principle :
1. At 80-85°C for 30 minutes for 3 successive days
2. In inspissator, the first exposure kills all the vegetative
forms, and in the intervals between the heating; the
remaining spores germinate into vegetative forms which
are then killed on subsequent heating;
Uses:-
1. Egg based media- e.g. Lowenstein-Jensen medium and
Dorset's egg medium.
2. Serum based media-e.g. Loeffler's serum slope.
 Hot air oven
Working principle:- l160 oC for 2 hours
Sterilization principle:- dry heat sterilization method
Uses:
1. For sterilization of glasswares- syringes, petridishes, flask,
pipettes and test tubes
2. Surgical instruments like scalpel, forceps, etc.
3. Chemicals, such as liquid paraffin, fats, glycerol, and glove
powder, etc
Precautions: it should not be overloaded, oven must be
allowed to cool before opening the doors since the glassware
may crack by sudden cooling
Sterilization control:
i. Biological indicator: Spores of nontoxigenic strains of
Clostridium tetani or Bacillus subtilis
ii. Thermocouples
iii. Browne's tube: contains a heat sensitive red dye which
turns green after being exposed to certiain temperature
 Incubator
Uses: for incubating culture plates and liquid media at
specified temperature (37oC) for growth of bacteria
 Candle jar

Procedure: a lighted candle is kept in air-tight container

loaded with inoculated culture plates. The candle is
extinguished by closing the container with its lid to
obtained the required atmosphere.
Uses:
1. For providing 5-10% carbon dioxide as required for
growing certain bacteria – H.influenzae
 Gas pack jar

Most commonly used method for anaerobiosis.

It uses a sachet containing sodium bicarbonate
and sodium borohydride which react chemically
in presence of water, to produce hydrogen and
CO 2 gas
Indicator of anaerobiosis:-
Chemical indicator: Reduced methylene blue
remains colorless in anaerobic conditions, but
turns blue on exposure to oxygen.
Biological indicator: Plate inoculated with
Pseudomonas is incubated along with other
inoculated plates for anaerobic culture. Absence
of growth of Pseudomonas ( which is an obligate
aerobe) indicates that perfect anaerobiosis
McIntosh & Filde’s Jar
For creating anaerobiosis.
Procedure:-
1. Evacuation of air and replacement with
hydrogen gas: done manually, by using a Kipp's
apparatus.
2. Removal of residual oxygen: this is done by
using a catalyst
Indicator of anaerobiosis:-
Chemical indicator : Reduced methylene blue remains
colorless in anaerobic conditions, but turns blue on
exposure to oxygen.
Biological indicator: Plate inoculated with
Pseudomonas is incubated along with other
inoculated plates for anaerobic culture. Absence of
growth of Pseudomonas ( which is an obligate
aerobe) indicates that perfect anaerobiosis
Tuberculin syringe
Uses:
1. In tuberculin test
2. Lepromin test
3. BCG vaccination
4. To inject a given dose of insulin
Sterilization:- glass syringe by hot air oven
Plastic syringe by gamma radiation