1. Instruments Techniques and principals
in Microbiology
1. Instruments used in microbiology are not
fully automated
2. Minimized human interference
3. To Aid in rapid diagnosis of infections
4. Minimized turnaround time
5. Safe and Hazard less enviornment

2. POLY STAINER 5300 automated
system for Gram stain
• The Polystainer is a fully
user programmable
robotic stainer capable
of delivering high
quality stained slides
for clinical and research
Microbiology labs
• The Polystainer will
take over the tedious
job of slide staining and
free lab staff to get on
with the work only
skilled scientists are
trained to do

3. Ittakes 5-10 minutes to stained the slides
Maximum 20 slides are stained

4. AUTOMATED BLOOD CULTURE
BACTEC™ 9240
• Need for Automated blood culture is,
• The detection of microorganisms in a patient’s blood
has diagnostic and prognostic importance. Blood
cultures are essential in the diagnosis and treatment
of the etiologic agents of sepsis.
• the expeditious detection and identification of blood
borne bacterial pathogens is an important function
of the diagnostic microbiology laboratory

5. • The BACTEC 9000 series of blood culture instruments are designed for
the rapid detection of microorganisms in clinical specimens.
• The sample to be tested is inoculated into the vial which is entered into
the BACTEC instrument for incubation and periodic reading.
• Each vial contains a sensor which responds to the concentration of CO2
produced by the metabolism of microorganisms or the consumption
oxygen needed for the growth of microorganisms.
• The sensor is monitored by the instrument every ten minutes for an
increase in its fluorescence, which is proportional to the increasing
amount of CO2 or the decreasing amount of O2 present in the vial.
• A positive reading indicates the presumptive presence of viable
microorganisms in the vial.
AUTOMATED BLOOD CULTURE
BACTEC™ 9240

6. • Media used for this instruments is as under
 BACTEC PLUS Aerobic/F Culture Vial
o 25 mL Enriched Soybean-Casein Digest broth (TSB)
o 0.05% Sodium Polyanetholesulfonate (SPS)
o Cationic and Non-ionic Adsorbing Resins
o Carbon dioxide (CO2) and Oxygen (O2)
o Sensor for the detection of fluorescence
 BACTEC™ PLUS Anaerobic/F Culture Vial
o 25 mL pre-reduced enriched Soybean-Casein Digest broth.
o 0.05% SPS
o Resins
o CO2 and Nitrogen gas (N2)
o Sensor for the detection of fluorescence.
 Volume: 3 to 10 mL of blood in each vial
 in paeds vial is 0.5 to 5 mL of blood is acceptable.
AUTOMATED BLOOD CULTURE
BACTEC™ 9240

7.  BACTEC PEDS PLUS/F Culture Vial
o 40 mL Enriched Soybean-Casein Digest broth
o 0.02% SPS
o Resins
o CO2 , O2
o Sensor for the detection of fluorescence
 BACTEC MYCO/F LYTIC
AUTOMATED BLOOD CULTURE
BACTEC™ 9240

10. MICROBIAL IDENTIFICATION USING
THE BIOMÉRIEUX VITEK® 2 SYSTEM
• The VITEK 2 is an automated microbiology system utilizing growth-based technology.
The system is available in three formats (VITEK 2 compact, VITEK 2, and VITEK 2 XL) that
differ in increasing levels of capacity and automation. Figure 1 shows the VITEK 2
compact system. All three systems accommodate the same colorimetric reagent cards
that are incubated and interpreted automatically.
• The reagent cards have 64 wells that can each contain an individual test substrate.
Substrates measure various metabolic activities such as
acidification, alkalinization, enzyme hydrolysis, and growth in the presence of inhibitory
substances.
• An optically clear film present on both sides of the card allows for the appropriate level
of oxygen transmission while maintaining a sealed vessel that prevents contact with the
organism-substrate admixtures.
• Each card has a pre-inserted transfer tube used for inoculation (described below). Cards
have bar codes that contain information on product type, lot number, expiration
date, and a unique identifier that can be linked to the sample either before or after
loading the card onto the system. Figure 2 shows the GN card.

12. Anaerobic Chamber
• Anaerobiasis is maintained in a gastight box or chamber by a
gas mixture containing or 80-90% Nitrogen(N2),5% Hydrogen
(H2), and 5-10% CO2 and by using a palladium catalyst.
Usually the system have a positive pressure inside the
chamber. The catalyst reduces the oxygen to water, thus
removing atmospheric oxygen from the chamber
• Carbon di oxide is included because many anaerobes require
it for growth
• Add anaerobic indicator to check the system how works well

14. CLINITEK 500
• The Clinitek 500 is a semi automated, bench top instrument
that performs urinalysis
• Testing using Siemens Multistix 10 SG reagent strips.
• The analyzer is a reflectance photometer that analyzes the
color and intensity of the light reflected from the reagent area
and displays the results in clinically meaningful units.
• No calculations are required.
• The Siemens Multistix 10 SG reagent strip includes testing
areas for glucose, bilirubin,ketones, specific
gravity, pH, protein, urobilinogen, nitrite, hemoglobin and
leukocyte esterase.

16. LabUMat- UriSed Urinalysis Analyzer
• The LabUMat test strip analyzer and UriSed microscopic sediment analyzer both
manufactured by 77 Elektronika
• Since all necessary measurements which have to be done on urine
samples analyze completed by this integrated system in one process,
combination of LabUMat and UriSed accelerates laboratory
throughput and provides the most effective and reliable solution for
complete and professional urine analysis.
• The LabUMat is an automated desktop instrument designed for high
volume urine test strip reading
• Automated test tube handling and test strip dipping
• The URISED is a professional urine sediment analyzer detecting urine particles in
urine sediment with high accuracy.
• This instrument performs sample preparation, takes several pictures of each
sample through its built-in microscope and evaluates them using the UriSed
Evaluation Module .
• which is a high quality image processing software

18. BACTEC™ MGIT™960
• The BACTEC™ MGIT™ 960 System is designed for the rapid
detection of mycobacteria in all types of clinical specimens
except blood and urine. (Each tube contains Modified
Middlebrook 7H9 Broth Base, Casein peptone )
• The system includes a liquid culture medium (BBL™ MGIT™
Mycobacteria Growth Indicator Tube), a growth supplement
and an antibiotic mixture (BBL™ MGIT™ PANTA™
• The BACTEC™ MGIT™ Growth Supplement provides
substances essential for the growth of mycobacteria.
• BBL MGIT PANTA contains a mixture of antimicrobial agents
used to suppress the growth of contaminating bacteria.

19. • A fluorescent compound is embedded in silicone on the bottom of
each of the BBL MGIT broth tubes. This compound is sensitive to
the presence of oxygen dissolved in the broth.
• Initially, the large amount of dissolved oxygen quenches the
emissions from the compound and little fluorescence can be
detected. Later, actively respiring microorganisms consume the
oxygen and allow the fluorescence to be detected.
• The BACTEC™ MGIT™ 960 System monitors the tubes for increasing
fluorescence. Analysis of the fluorescence is used to determine if
the tube is instrument-positive; i.e., the test sample contains viable
organisms. Culture tubes which remain negative for a minimum of
42 days (up to 56 days) and which show no visible signs of positivity
are removed from the instrument as negatives.
BACTEC™ MGIT™960

21. Fluorescence microscope
• A fluorescence microscope is an optical microscope used to
study properties of organic or inorganic substances using the
phenomena of fluorescence instead of, or in addition to,
reflection and absorption.[
• In fluorescence microscopy, the sample you want to study is
itself the light source. The technique is used to study
specimens, which can be made to fluoresce.
• The fluorescence microscope is based on the phenomenon
that certain material emits energy detectable as visible light
when irradiated with the light of a specific wavelength.

22. Fluorescence microscope
• The sample can either be treated with fluorescing chemicals.
• The Sample Gets Excited
The basic task of the fluorescence microscope is to let
excitation light radiate the specimen and then sort out the
much weaker emitted light to make up the image. First, the
microscope has a filter that only lets through radiation with
the desired wavelength that matches your fluorescing
material. The radiation collides with the atoms in your
specimen and electrons are excited to a higher energy level.
When they relax to a lower level, they emit light.

23. Fluorescence microscope
• To become visible, the emitted light is separated from the
much brighter excitation light in a second filter. Here, the fact
that the emitted light is of lower energy and has a longer
wavelength is used. The fluorescing areas can be observed in
the microscope and shine out against a dark background with
high contrast.
• Fluorescence microscopy is a rapid expanding technique, both
in the medical and biological sciences. The technique has
made it possible to identify cells and cellular components with
a high degree of specificity.

25. Principle of Fluorescence
1. Energy is absorbed by the atom which
becomes excited.
2. The electron jumps to a higher energy level.
3. Soon, the electron drops back to the ground
state, emitting a photon (or a packet of light) -
the atom is fluorescing.

27. Biosafety cabinet
• A biosafety cabinet (BSC), biological safety cabinet, or
microbiological safety cabinet is an enclosed, ventilated
workspace for safely working with materials contaminated
with (or potentially contaminated with) pathogens in the
laboratory. Several different types exist, differentiated by the
specifics of construction.
• The primary purpose of a BSC is to serve as the primary
means to protect the laboratory worker and the surrounding
environment from pathogens. All exhaust air is HEPA-filtered
as it exits the biosafety cabinet, removing harmful bacteria
and viruses. This is in contrast to a laminar flow clean
bench, which blows unfiltered exhaust air towards the user
and is not safe for work with pathogenic agents.[

28. Biosafety cabinet
• The U.S. Centers for Disease Control and Prevention (CDC)
classifies BSCs into three classes. These classes and the types
of BSCs within them are distinguished in two ways: the level
of personnel and environmental protection provided and the
level of product protection provided.
• Class I cabinets provide personnel and environmental
protection but no product protection
• Class II cabinets provide both kinds of protection (of the
samples and of the environment) since makeup air is also
HEPA-filtered.