PRESENTATION ON

PROJECT REPORT
PRACTICAL IN
BIOCHEMISTRY, PATHOLOGY AND MICROBIOLOGY

DONE BY [III BSC. BBC]-

HIMANI MISHRA (16AD697)
SPANDANA ALLU (16AD727)
SURABHI KATIYAR (16AD740)
 CONTENT
• INTRODUCTION
• SAMPLE COLLECTING TUBES
• SPECTROPHOTOMETER
• FULLY AUTOMATED BIOCHEMISTRY ANALYZER
• Hb ELECTROPHORESIS
• ELISA READER
• ELECTROLYTE ANALYZER
• CELL COUNTER
• FULLY AUTOMATED IMMUNOASSAY ANALYZER
• IMMUNOASSAY ANALYZER(Cobas e 411 Roche)
• FULLY AUTOMATED BACTERIAL IDENTIFICATION(VITEK 2)
• AUTOMATIC TISSUE PROCESSOR
 INTRODUCTION
• Clinical chemistry (also known as chemical pathology, clinical
biochemistry or medical biochemistry) is the area of chemistry that is
generally concerned with analysis of bodily fluids for diagnostic and
therapeutic purposes. It is an applied form of biochemistry.

• Most current laboratories are now highly automated to accommodate

the high workload typical of a hospital laboratory. Tests performed are
closely monitored and quality controlled.

• All biochemical tests come under chemical pathology. These are

performed on any kind of body fluid, but mostly on serum or plasma.
 SAMPLE COLLECTING TUBES
Pathology specimens should meet defined standards of patient identification and specimen labelling
to ensure that the chain of information custody from patient to sample to laboratory to report is
unbroken.
Different types of sample collecting tubes –
ADDITIVE CAP COLOUR BLOOD VOLUME SUITABLE FOR

Most chemistry, including drug levels and

serological tests that require serum. Also for
Clot Activator,
RED 6mL immunohaematological tests that require clotted
Plain
blood such as abnormal blood group antibody
tests and cross-matching requests.

Most haematological tests, HbA1C,

K2EDTA (Di- Cyclosporine, Homocysteine, and molecular
Potassium Ethylen genetics tests (when using blood DNA). After the
PURPLE 3mL
e Diamine- tube has been filled with blood drawn by
Tetracetate) vacuum, it should be gently inverted at least 6 to
10 times to prevent clotting.
 Tests requiring plasma or whole blood (such as cytogenetics
when using blood DNA) and STAT biochemistry tests such as
Lithium
GREEN 4mL electrolytes, renal screen, ammonia test. After the tube has
Heparin
been filled with blood drawn by vacuum, it should be
inverted gently at least 6 times to prevent clotting.
Coagulation studies. The ratio of blood to anticoagulant is
critical for valid prothrombin time and activated partial
thromboplastin time results. Allow 2.7mL of blood to be
Sodium Citrate BLUE 2.7mL
drawn by vacuum. This tube should be inverted gently at
least 3-4 times in order to prevent clotting. (Must fill up to
the mark)

Sodium
Glucose tests. After the tube has been filled with blood drawn
Fluoride
GREY 6mL by vacuum, it should be inverted gently at least 6 times to
/ Potassium
prevent clotting.
Oxalate

SST II, Clot

All tests requiring serum except those few that need red cells
activator &
as well (such as abnormal blood group antibody screen, cold
serum YELLOW 5mL
agglutinins). After centrifugation, the gel forms an effective
gel separator,
barrier between the blood clot and the serum.
Plain
SPECTROPHOTOMETER
The spectrophotometer is a routinely used instrument in scientific research.
Spectrophotometry is the quantitative measurement of how much a chemical
substance absorbs light by passing a beam of light through the sample using a
spectrophotometer.
A spectrophotometer is commonly used for the measurement of transmittance or
reflectance of solutions, transparent or opaque solids, such as polished glass, or
gases.
PRINCIPLE - each compound absorbs or transmits light over a certain range of
wavelength.
Spectrophotometer is important equipment used in many biochemical
experiments that involve DNA, RNA, and protein isolation, enzyme kinetics and
biochemical analyses. The spectrophotometer is used to measure colored
compounds in the visible region of light (between 350 nm and 800 nm), thus it
can be used to find more information about the substance being studied. In
biochemical experiments, a chemical and/or physical property is chosen and the
procedure that is used is specific to that property in order to derive more
information about the sample, such as the quantity, purity, enzyme activity, etc.
This UV-VIS Spectrophotometer can be used extensively for qualitative and
quantitative analysis in such field as biochemical research and industry,
pharmaceutical analysis and production, education, environmental protection,
food industry, clinical examination, sanitation and antiepidemic etc.
FULLY AUTOMATED BIOCHEMISTRY ANALYZER
Fully automatic biochemistry analyzer (FABCA) is a high performance micro-controller based Photometric
biochemistry analyzer used to measure various blood biochemical parameters such as blood glucose, urea, protein,
and bilirubin etc. that are associated with various disorders such as diabetes, kidney diseases, liver malfunctions and
other metabolic derangement’s. This equipment is helpful in diagnosing health disorder. Automatic biochemistry
analyzer (ABC) analyzer facilitate to be controlled via laptop or PC by using the interfacing/front end software or as
standalone unit.
PRINCIPLE - Photometry
SPECIAL FEATURES –
Throughput: Double Reagent: 200 Tests/hour
Single Reagent: 260 Tests/hour
• Random Access with STAT function
• Assay Modes: End Point, Fixed time (2-point), Kinetic Rate-A, Kinetic
Rate-B
• Reagent Positions: 30 positions for 20 ml bottles
• 60 dismountable, easily replaceable and reusable cuvettes
• 8 wavelengths: 340, 405, 505, 546, 578, 600, 670, 700 nm
• Twin Laundry System, 7-step automatic washing
Sample Positions: Maximum 30 positions with 15 positions flexible for reagent in sample tubes.
Calibration Points: K-Factor, Linear (one, two and multi point), Logic-log, Spleen, Exponential, Polynomial (second,
third and fourth order) Multipoint curves for up to 6 points.
• Low water consumption of 4 litres / hour
• Easy-to-use Windows based software (XP 2007)
Innovative Feature
It is a fully automated, random access, patient prioritized analyzer with through-
put of 200 tests / hour for double reagent and 260 tests / hour for single
reagent, with on board ISE module , making it ideal for small to mid-sized
laboratory.
Enhanced Lab Efficiency
It is equipped with sample and reagent probe with liquid level sensor. Analyzer
is programmed with internal and external probe washing.
Reaction Disk Assembly
It is provided with 60 dismountable, easily replaceable and reusable cuvettes.
Analyzer is equipped with twin laundry and 7- step automatic washing system,
with low water consumption of 4 litres / hour.
 HB ELECTROPHORESIS
A haemoglobin electrophoresis is a test performed to measure the types of
hemoglobin present in bloodstream. Haemoglobin is a protein present in
Red blood cells and helps carry oxygen and supply it to the cells.
Different haemoglobin types have different electrical charges, and this
Provides the base for the haemoglobin electrophoresis process. The
process involves passing an electrical current through the haemoglobin in a
blood sample to separate different haemoglobin types. The separation occurs at different rates for different types of
haemoglobin. These types that form bands can be compared with the pattern found in a normal blood sample.
Haemoglobin electrophoresis at pH 8.4–8.6 using cellulose acetate membrane is simple, reliable and rapid. It is
satisfactory for the detection of most common, clinically important haemoglobin variants.
PRINCIPLE - At alkaline pH, haemoglobin is a negatively charged protein, and when subjected to electrophoresis will
migrate toward the anode (+). Structural variants that have a change in the charge on the surface of the molecule at
alkaline pH will separate from haemoglobin A. Haemoglobin variants that have an amino acid substitution that is
internally sited may not separate, and those that have an amino acid substitution that has no effect on overall charge
will not separate by electrophoresis.
ADVANTAGE –
Improved Diagnosis - When doctors suspect a patient has a genetic blood disorder, such as sickle cell
anemia, haemoglobin electrophoresis separates the haemoglobin types for examination. This helps to
identify abnormal haemoglobin. One characteristic of sickle cell anemia is a specific type of
haemoglobin, and this is true with many blood disorders. The separation allows technicians to locate
each blood factor and make a diagnosis.
Simplicity - Electrophoresis is a fast and easy technique. The improvements in this field enable
technicians in a laboratory to perform electrophoresis will little effort. Modern labs use gel solutions as
the conductor for the electrical charge. This benefits everyone, because the separation is quick and easy
once the specimen becomes available.
Low-Cost Material - The material necessary to perform electrophoresis costs little and is easy to prepare.
According to Molecular Station, the common media for electrophoresis, gel, is straightforward to mix
and pour. Supply cost is one factor in the expense of laboratory testing.
Reliability - Of all the benefits electrophoresis offers, reliability is the most essential. The technique is
fast, simple and it works. If done correctly, electrophoresis will separate all molecules in a specimen. For
a technician, the ability to rely on a technique makes their job easier and allows for effective results.
DNA testing and analysis is a complicated process, but one that allows for forensic identifications,
species analysis and diagnosis of disease. Electrophoresis helps make that happen.
HAEMOGLOBIN ELECTROPHORESIS PATTERNS IN
BETA THALASSEMIA PATIENT
The following list corresponds to this image of an alkaline hemoglobin
electrophoresis -
i) Lanes 1 and 2: normal patient specimen
Hb A is over 98% with a small amount of Hb A2 visible
ii) Lanes 3 and 4: Beta thalassemia minor
Hb A is decreased to 94%, Hb A2 is increased at 5%, and Hb F is 1%
iii) Lanes 5 and 6: Delta-beta thalassemia major
No Hb A or A2 is present, Hb F is 100%
iv) Lanes 7 & 8: Delta-beta thalassemia intermedia
Hb A is 8.5%, Hb A2 is 3.5% and Hb F is 88%
v) Lane 9: AF control
vi) Lane 10: ASC control
NOTE - AF and ASC are labels and do not indicate the order of migration
 ELISA READER
The enzyme-linked immunosorbent assay, also known by the acronym, ELISA, was created in the 1970s. This common lab test
measures the concentration of an analyte, which are generally antibodies or antigens in a particular solution. With ELISA,
quantitative results can be detected, which sets it apart from other similar type tests.
There are four different types of ELISA tests:
• Direct: This method is the fastest and has fewer steps. It is also less prone to an error.
• Indirect: This method has increased sensitivity and it costs less as fewer labeled antibodies are
required.
• Sandwich ELISA: With this method, there are more steps involved. However, the results are highly
specific.
• Competition or Inhibition ELISA: This method is usually used when only one antibody is
available or when the analyte is small.
ELISAs are typically performed in 96-well (or 384-well) polystyrene plates, which passively bind anti-
bodies and proteins. It is this binding and immobilization of reagents that makes ELISAs so easy to
design and perform. Having the reactants of the ELISA immobilized to the microplate surface makes it
easy to separate bound from non-bound material during the assay. This ability to wash away
nonspecifically bound materials makes the ELISA a powerful tool for measuring specific analytes within a crude preparation.
A detection enzyme or other tag can be linked directly to the primary antibody or introduced through a secondary
antibody that recognizes the primary antibody. It can also be linked to a protein such as streptavidin if the primary
antibody is biotin labeled. The most commonly used enzyme labels are horseradish peroxidase (HRP) and alkaline
phosphatase (AP). Other enzymes have been used as well, but they have not gained widespread acceptance because
of limited substrate options. These include β-galactosidase, acetylcholinesterase and catalase. A large selection of
substrates is available for performing ELISA with an HRP or AP conjugate. The choice of substrate depends upon the
required assay sensitivity and the instrumentation available for signal-detection (spectrophotometer, fluorometer or
luminometer).

Comparison of direct and indirect ELISA detection methods

Direct ELISA detection

•Quick because only one antibody and fewer steps are used.
Advantages
•Cross-reactivity of secondary antibody is eliminated.

Immunoreactivity of the primary antibody might be adversely affected by labeling

with enzymes or tags.
Labeling primary antibodies for each specific ELISA system is time-consuming and
Disadvantages
expensive.
No flexibility in choice of primary antibody label from one experiment to another.
Minimal signal amplification.
Indirect ELISA detection
•A wide variety of labeled secondary antibodies are available commercially.
•Versatile because many primary antibodies can be made in one species and
the same labeled secondary antibody can be used for detection.
•Maximum immunoreactivity of the primary antibody is retained because it is
Advantages not labeled.
•Sensitivity is increased because each primary antibody contains several
epitopes that can be bound by the labeled secondary antibody, allowing for
signal amplification.
•Different visualization markers can be used with the same primary antibody.
•Cross-reactivity might occur with the secondary antibody, resulting in
Disadvantages nonspecific signal.
•An extra incubation step is required in the procedure.
 ELECECTROLYTE ANALYZER
Pro Electrolyte Analyzer is completely automated, microprocessor controlled electrolyte system that uses current ISE
technology to make electrolyte measurements. It measures various combinations of sodium, potassium, Ionized
Calcium, Lithium, pH and Chloride in whole blood, serum, plasma, and urine.
PRINCIPLE-direct measurement with ion selective electrode (ISE)
FEATURES -
• Multilingual support.
• Levey-Jennings chart for NABL and seamless integration to LIS(Lab information system).
• 7" inch high definition LCD with capacitive touch display.
• Excellent precision and reliability.
• Aspirate 1 sample and acquire 6 different Critical Care results. (Na, K, iCa, pH, Cl Li)
• Integrated Parameter conversion functionality.
• One single Reagent Pack with all 10 combinations.
• Numeric alpha numeric input option with 15 digit operator patient ID.
• External barcode scanner, mouse keypad interfacing option.
• Self-calibration once new reagent pack is installed.
• Self-probe wiping facility with 3 different aspiration modes
(sample tube, sample syringe capillary).
• Maximum 2,00,000+ sample storage capacity.
• The result can be recalled by Date, Patient ID, Patient name,
by parameters etc.
• Extremely low cost per test.
• Two USB output ports. (USB 2.0)
• Optional battery backup much more.
• Sample type – whole blood, serum, plasma, CSF and diluted urine.
• Sample volume – 140 μl
• Reading time – 65 seconds
• Speed – 55 samples per hour.
 CELL COUNTER
Automated cell counters are machines that automatically count cells. The sample is loaded into an automated cell
counter and it is forced through a small tube while the automated cell counter uses optical or electrical impedance
sensors to count how many cells go through the tube. Used in medical and research labs, automated cell counters can
be used on blood or urine samples to determine the number and types of cells present or to check the viability of a
cultured cell line for research purposes.
Cell counter uses microscopy with auto-focus that analyzes multiple focal planes to identify the best plane. Without
requiring any user input, the sophisticated cell counting algorithm uses the image acquired from the best focal plane to
identify cells and exclude debris, thereby calculating the total cell count. The auto-focus leads to highly reproducible
cell counts with reduced user-to-user variability compared to a haemocytometer and cell counters with manual focus.
Using auto-focus instead of subjective manual focusing is especially important when assessing cell viability because an
incorrectly selected focal plane will lead to inaccurate results.
Key Features and Benefits
• Compatible with a broad range of cell sizes and types — counts cell lines, primary cells (from tissue or blood), and
stem cells.
• Innovative auto-focus technology — removes the variation associated with manual focusing and leads to precise
cell counts in 30 seconds.
• Cell size gates — user selects a population of interest in complex samples, such as primary cells, or lets the cell
counting algorithm do all the work.
• Cell viability — analyzes cells accurately using multifocal plane analysis.
FEATURES:
• Throughput: Up to 60 samples/hour
• Reagents: Only 4 onboard reagents and 1 diluent
• Basophils counted through specific channel
• High resolution matrix includes the determination of 2 subpopulations: Atypical
• Lymphocytes (ALY) and Large Immature Cells (LIC)
• Data management on stand-alone PC.
• Micro-sampling from whole blood (CBC : 30 µL - DIFF: 53 µL)
• Cytochemistry, Impedance (real cell volume measurement) & Optical (analysis of the
internal cellular structure by measuring light absorbance) - DHSS(Double Hydrodynamic
Sequential System) Technology.
• Perfect homogenization of blood samples with reagents – MDSS (Multi-Distribution
Sampling System)Technology.
• Power supply – 100 to 240V (± 10%)50 to 60 Hz.
• Throughput- 60 tests/hour.
• Memory- 10,000 results + Graphics
 FULLY AUTOMATED IMMUNOASSAY ANALYZER
An immunoassay is a biochemical test that measures the presence or concentration of a macromolecule or a small
molecule in a solution through the use of an antibody (usually) or an antigen (sometimes).An immunoassay analyzer
is used in hospital and clinical laboratories to run automated biochemical tests to detect the presence and
concentration of substances in the samples.
PRINCIPLE-In the presence of complimentary antigen and antibody, the paratope of the antibody binds to the
epitope of the antigen to form an antigen-antibody or an immune complex. Estimating the levels of such immune
complex by use of labelled antibodies form the basis of CLIA. It involves use of stationary solid particles coated
either with the antigen or antibody of interest. Post incubation, which ensures intact immune complexes are formed,
substrate is added. This results in generation of light, the intensity of which is directly proportional to the amount of
labelled complexes present and which indirectly aids in quantification of the analyte of interest. The intensity of light
is measured in terms of Relative Light Units (RLU).
• ASSAYS PERFORMED- Thyroid, Fertility, Tumor Marker, Inflammation, Prenatal Screening, Anemia, Kidney
Function, Hepatic Fibrosis, Bone Metabolism, Diabetes, Cardiac Marker, Allergy, TORCH, Infectious Diseases.
• TECHNOLOGY-Magnetic Nanoparticles Based Enhanced Immuno chemiluminescence
• SAMPLE REQUIRED- Serum, Plasma, Urine, Whole Blood
• SAMPLE VOLUME- 10-100
• SAMPLES ONBOARD- 50
• REAGENT SYSTEM- Closed
FEATURES:
• Clot & Bubble Detection
• Automatic Dilution
• Nano-magnetic microbeads separation phase
• Throughput: Maximum 180 tests/hr
• Sample and reagent continuous loading
• On board capability: up to 40 samples
• Random access or batch mode, STAT
• 2-point calibration with 10 point preset master curve.
• Refrigerated reagent area
• Reagent recognition: RFID reader
• Level sensor and clot detection
• Auto dilution for higher titer samples
The main advantage of this technology includes sensitivity and its ability to be unaffected by background signals.
Also, the analyzers working under this principle are simple in design and operation. CLIA (chemiluminescence
immunoassay analyzer) uses two important technologies, one is labeling technology which determines reaction
mode; and the other is separation technology which determines the sensitivity, accuracy and precision of the
reagents.
 IMMUNOASSAY ANALYZER (COBAS E 411 ROCHE)
PRINCIPLE- Photometry
Features-
Bench top analyzer for heterogeneous immunoassays. Cobas e411 offers rapid STAT and turnaround time, an on-
board Capacity of 18 tests and throughput of up to 88 tests per hour. Sample carrier options include disc or
Roche/Hitachi five-position rack.
Benefits-
• Easy to operate - The customized keyboard and easy-to-learn software make training and operation simple and
keep user involvement to a minimum.
• Unique programming-by-loading concept - Barcode-based data entry is carried out automatically by loading
reagents, controls and calibrators onto the system – a rapid, robust and safe procedure.
• STAT facilities for urgent samples - Cobas e 411 disk system features two STAT sample positions that can be
accessed at any time, delivering results rapidly in response to clinicians‘ requests. The cobas e 411 rack system
features a STAT port for immediate access emergency testing.
• Innovative technology - Novel Electrochemiluminescence (ECL) technology provides superior
analyticalperformance.Increasedsensitivitymeansthatextremelylowlevelsofantigen,as well as subtle changes in levels
can be detected.
 cobas e 411 analyzer Specifications-
 System-Fully automated, random access system for immunoassay analysis.It is available as both, a disk system and a
rack system.
 System components-Analytical module including Window XP embedded operated touch screen PC. Sample
handling module: rack or disk operated
 Sample throughput-Up to 88 samples/hr (theoretical max)
 Test throughput- Up to 88 tests/hr (theoretical max)
 Number of channels- 18 channels/reagent slots for up to 18 different assays
 Programmable parameters- Max 60 assays definable via 2D-barcode (programming by loading)
 Sample types- Serum, Plasma, Urine
 Sample input/output-
• Load/unload capacity-30 samples (disk);75 samples on 15 racks
• Rack- RD standard 5 position rack
• Rack types-Routine, STAT, Control, Calibrator
• STAT handling-Any unoccupied position on the sample disk, dedicated STAT port on rack feeder.
 Sample container types-
• Primary tubes- 5 –10 ml; 16 x100, 16 x 75, 13 x100, 13 x 75mm
• Sample cup- 2, 5 ml
 Water/waste requirements-
• Water container: 3 Litres
• Water requirements: 10 μS/cm or
• 0.1 mega Ohm, bacteria-free
• Water consumption: approx. 3 L for 250 tests; approx. 12 mL/cycle
 Regulatory requirements- GS, CE, UL, C-UL, CB-report and certificate
 Operating conditions-
• Ambient temperature: 18 to 32 °C (64.4 °F to 89.6 °F)
• Ambient humidity: 20 % to 80 %
• Noise Output: 60 dbA (stand-by mode)
• 63 dbA (avg. during operation)
 Physical dimensions-
• Width: 1200 to 1700 mm (disk/rack)
• Depth: 730 to 950 mm (disk/rack)
• Height: 560 mm (w/o PC unit)
 Weight- Approx. 170 kg (disk) and 210 kg (rack)
 FULLY-AUTOMATED BACTERIAL IDENTIFICATION
(VITEK 2)
Fully automated microbial identification system has everything healthcare laboratories need for fast, accurate
microbial identification, and antibiotic susceptibility testing. The innovative microbial identification system includes an
expanded identification database, the most automated platform available, rapid results, improved confidence, with
minimal training time.
The fully automated bacterial identification system provides greater automation while increasing safety and
eliminating repetitive manual operations. The rapid response time means results can be provided more quickly than
with the manual microbial identification techniques.
This automated system offers-
• Intuitive software
• User interface screen for immediate notification of system status to increase productivity.
• Unique vacuum filler provides both safety and the highest level of automation.
• Elimination of many manual steps:
• Designed for simple temperature verification.
• Completed tests are automatically ejected into an ergonomic trash container.
This system features - Three-step setup procedure; High level of automation; Intuitive, icon-driven software; Familiar
Windows format; Easy graphical representation of phenotypes; One-click result validation.
USES-
• Microbial Identification - bacteria and yeast identification (ID).
• Antibiotic susceptibility testing (AST) and resistance mechanism detection.
• Epidemiologic trending and reporting.
FEATURES AND BENEFITS
• Reduce time to microbial identification and antibiotic susceptibility testing results.
• Offer an extensive identification and susceptibility menu.
• VITEK 2 technology with the ADVANCED EXPERT SYSTEM offers:
• Knowledge base developed from >100,000 references.
• >2,000 described phenotypes.
• >20,000 MIC distributions.
• >100 resistance mechanisms detected.
• >99 organisms.
• On average, provides a resulting range of five to seven MIC doubling dilutions per antibiotic.
• Extended MIC range to enable low-level resistance detection.
• Resistance-oriented results that highlight unusual phenotypes.
• Deduced antibiotic results to meet formulary requirements.
 ANTIBIOTIC SUSCEPTIBILITY TESTING (AST)
• Antibiotic susceptibility testing (AST)is essential to adapt the patient’s antibiotic treatment and fight antibiotic
resistance.
• The Minimum Inhibitory Concentration (MIC) is a measure of the sensitivity of microbes to antibiotics or
antifungals. Organisms are tested for growth in different concentrations of antibiotics. This allows microbiologists
to see which antibiotic concentration (MIC) will be effective against the disease agent.
OBJECTIVE - The test determines the susceptibility of microbial species against different antibiotic agents.
PRINCIPLE - The introduction of various antimicrobials for treating
variety of infections showed the necessity of performing antimicrobial
testing as a routine procedure in all microbiology laboratories. In
laboratories it can be made available by using antibiotic disc which
will diffuse slowly into the medium where the suspected organism is
grown. The basic principle of antibiotic susceptibility testing has
been used in microbiology laboratories over 80 years. Various
chemical agents such as antiseptics, disinfectants, and antibiotics are
employed to combat with the microbial growth. Antimicrobial susceptibility tests (ASTs) basically measures the ability
of an antibiotic or other antimicrobial agent to inhibit the invitro microbial growth.
 HISTOPATHOLOGY
The word 'histopathology' is derived from a combination of three Greek words: histos meaning tissue, pathos
meaning disease or suffering, and logos which refers to study in this context
• Histopathology is the examination of tissues from the body under a microscope to spot the signs and characteristics
of disease. Histology is the study of tissues, and pathology is the study of disease.
• So taken together histopathology literally means the study of tissues as relates to disease. A histopathology report
describes the tissue that has been sent for examination and the features of what the cancer looks like under the
microscope. A histopathology report is sometimes called a biopsy report or a pathology report.
TYPES OF SAMPLES USED-

BIOPSY FNAC AUTOPSY AMPUTED LBC

LIMBS
Uterus Lump From dead body From alive person Vaginal discharge
Placenta Bump Upper limb Fluids
Salivary gland Lower limb
Cervical
Kidney
Uses of Histopathology
• investigate crimes e.g. look for causes of injury or death such as evidence of tissue damage by poisons, drugs or
possibly deliberately targeted biological pathogens - Forensic Pathology or Forensic Histopathology.
• Investigate historical artefacts containing biological tissue in sufficiently good condition to learn about the health
of long-deceased individuals - which can involve various types of histology and histopathology, e.g. bone
histology, dental histology and so on. This is similar to the
• Study of ancient diseases (an area of study known as 'paleopathology', sometimes spelt 'palaeopathology') using
histological techniques - Histology in Palaeopathology, or Histopalaeopathology.
Abstract
• The rodent hippocampal slice preparation is perhaps the most broadly used tool for investigating mammalian
synaptic function and plasticity. The hippocampus can be extracted quickly and easily from rats and mice and slices
remain viable for hours in oxygenated artificial cerebrospinal fluid.
• Moreover, basic electrophysisologic techniques are easily applied to the investigation of synaptic function in
hippocampal slices and have provided some of the best biomarkers for cognitive impairments. The hippocampal
slice is especially popular for the study of synaptic plasticity mechanisms involved in learning and memory.
Changes in the induction of long-term potentiation and depression (LTP and LTD) of synaptic efficacy in
hippocampal slices (or lack thereof) are frequently used to describe the neurologic phenotype of cognitively-
impaired animals and/or to evaluate the mechanism of action of nootropic compounds. All types of sectioning is
possible including sectioning for visual patching of neurological tissue, heart, and lung, and much more.
FEATURES
• Includes a Z-axis calibration unit
• Minimal Z-axis deflection (less than 1µm) at all speeds and amplitudes
• Z-axis blade adjust minimizer
• Blade holder angle to user requirement
• Set start and stop position of blade travel
• Tissue sample automatically retracted before blade returns to start point
• Full range of adjustable parameters with 8 customizable user profiles
• Vibration speeds from 50 to 120Hz
• Amplitudes from 0.5mm to 2.5mm
• Controlled blade advance at 10 microns per sec
• "Auto" programming by storage of the first slicing
• Ice water bath easily removed for cleaning
• Optional LED light and scope for clear observation
• Leaf spring vibratory mechanism for optimal longevity and accuracy
 AUTOMATIC TISSUE PROCESSOR
Automatic Tissue Processor is a compact and versatile equipment for staining process automization from stage one of
fixation of tissue and dehydration process to paraffin wax saturation. Glass door with locking arrangement provides
easy viewing and safety from tempering. Continuous agitation is provided by rotating S.S. Tissue basket to make
process more effective. A superior thermostat maintains desired temperature of wax bath.
This model is specially designed and immaculately profiled. It effectively
combines aesthetic appeal with functional utility. Most effective feature of the machine is that
only Basket Rotor along with the single bakelite beaker cover lifts up to change the
position on to the next stage of the processing cycle. All the other beakers remain
covered during each change of stage, thus keeping the loss of alcohol/reagents to a
minimal level due to evaporation.
Histopathological Instrument and equipment’s:- Microtome, Microtome knife, Timer, Hot air oven, Forceps, Scalpel,
dissecting set, Tissue floatation bath, Equipment for embedding and vacuum, Containers for holding specimens.
Steps for the Tissue processing - Grossing, Labeling, Fixation, Dehydration, Clearing, Impregnation, Embedding,
Microtomy, Staining, Microscopic observation .

Grossing Labeling Fixation Dehydration Clearing Impregnation

Tissue cuts Every tissue It prevent Water is Alcohol It remove the clearing
into small
pieces like need to from natural removed by Is removed reagent .paraffin. Tissue
3-4 mm give an decompose different grade by Xylene. transfer into P. Wax Ist,
identity for Exp... of Iso propyl IInd and IIIrd changes.
Recognize Formalin. It used
alcohol. Temperature should be 2-3
with two
50% Alcohol
changes I,& degree more than its
70%,90%,and IInd melting point
95%
Paraffin wax.
 CONCLUSION

• On the whole , This internship was a useful experience. We have gained new knowledge and skills.
The friendly welcoming of staff at Path labs made us enthusiastic to know more information. We got
in sighted into professional practices currently advocated in the field of biochemistry. We learned the
different facets of working within a well established laboratories.
• One of the fascinations are studying about various bioinstruments and working on it and also to
know that a fully automated analyzer is much capable than us to give out accurate results. In the
field of biochemistry, it is importance to have hands on experience and practical skills to understand
the concept.
• Over all, my internship was good to find out my strengths and weaknesses and to gain much
knowledge. It actually inspired me to know more about the world of biology.
THANK YOU