Instruments
1. LP needle (Lumbar puncture Needle)
1. LP needle (Lumbar puncture Needle)
• Is a long slender needle of 10 cm length and 1 to 1.5mm bore. Also has a
stillette
• Used to tap CSF
• Indications for LP(Lumbar puncture)
For diagnosis of type of meningitis (Viral or Tuberculous or bacterial/Fungal etc);
subarachnoid hemorrhage; Demyelinating diseases, blast cells in leukemia
For introduction of drugs ,anaestheic agents, contrast media
• Contraindications for LP:
Increased intracranial pressure, Infections in local area
• Sites of puncture: Lumbar-Between L3 &L4 vertebrae or L4-L5
Rarely: Cisternal or Ventricular puncture, Anterior fontanelle in infants
CSF analysis
• CSF opening pressure is measured using manometer.
Normal CSF pressure is 70 to 180mm of water in lying position
• CSF is collected in 3 sterile tubes and tested for the following
• 1st tube: Bacteriological examination-Grams stain , AFB stain, Culture etc
• 2nd tube:Biochemical tests(Sugar,protein,chlorides)
Decreased glucose levels are seen in Bacterial meningitis, and increased glucose in Viral
meningitis ; Chlorides are grossly increased in Tuberculous meningitis ;Proteins are highly
increased in Bacterial meningitis
• 3rd tube cell count and cytological examination (Neutrophils are seen in Bacterial
meningitis while in Viral and Tuberculous , it is mainly Lymphocytes),Blast cells in ALL
Normal CSF is clear, colourless, has mainly few lymphocytes( <5cells/cu mm )
CSF appears blood stained in subarachnoid haemorrhage/ traumatic tap,
(In traumatic tap,the CSF clears in subsequent tubes)
cloudy in Pyogenic and clear in Viral meningitis
Xanthochromic- pale Pink to Orange to Yellow colour of CSF –in supernatant of centrifuged CSF,
that happens in subarachnoid haemorrhage. Blood lysis in CSF liberating Hb which is slowly
converted to bilirubin
Lumbar Puncture
• Sites
• Vol of CSF collected
• CSF pressure measurement
• Tubes
• Tests done –Appearance, cob web coagulam
-Biochemical, Proteins, Glucose, Chloride
-Cells – Neutrophils, lymphocytes, malignant cells-
Blasts, CNS malig,other malig-breast,
- Gms stain,AFB stain,India Ink preparation
- Culture
- serological test-syphilis, Ig G and Ig M antibodies
2. Jamshidi’s bone marrow aspiration
and biopsy needle
• Jamshidi Bone marrow aspiration and biopsy needle is used. Only
aspiration means , it can be performed with Salah’s needle or Klimas
needle
• Indications for Bone marrow study: Megaloblastic anemia, Aplastic
anemia, Leukemias, ITP, Myelofibrosis; Parasitic diseases like
Leishmaniasis,Malaria; Storage disorders like Gaucher’s , Niemann
Pick ; Lymphoma staging, etc
• Contra indications: Coagulation factor disorders, Infection at the site
• Site of puncture: Posterior Iliac crest in adults, Medial surface of tibia
in children. Other sites : spinous process of vertebrae,Sternum opp to
2nd intercostal space (aspiration only)
[Link] biopsy needle
Trucut biopsy needle
• This helps us to collect good patient samples with
minimal trauma to patients
• This is used to take biopsy of superficial as well as
deep seated lesions. For deep lesions, the biopsy
is taken with USG or CT guidance
• Once the needle is inside the lesion(with or
without guidance), then the gun is clicked and the
cutting needle inside takes a core of tissue
• Tissue thus taken is immediately fixed in 10 %
buffered formalin.
• Trucut biopsy needle can be used for Liver biopsy
also
• Indications for liver biopsy: Cirrhosis, Primary or
Secondary liver tumours; PUO; storage disorders
like Glycogen storage disorders, Wilson’s disease;
Diagnosis of Granulomas-TB,Fungal ;
• Contraindications: Coagulation disorders: When
Prothrombin time is >3; Platelet count is <80,000
cells/cu mm; Hydatid cyst of liver ; Hemangioma
of Liver
• Site Commonly at 8th or 9th intercostal space
4. Ayre’s spatula
• Made of soft wood
• Used to collect Cervical, Vaginal ,Buccal smears for
cytological study
Cytology
• Cytology is the study of cells that have been
exfoliated freely from the tissues or collected by
brushing, scraping, washing or by needle
aspiration(Fine Needle Aspiration Cytology)
• FNAC is the study of cellular sample obtained
through a fine needle under negative pressure
• Fixative used for cytological study is Isopropyl
alcohol
• Wet fixed smears stained with Papanicolaou stain
• Air dried smears are stained with MGG stain
Cytology- Advantages
:Easy to perform & gives earlier diagnosis.
Relatively painless,gives speedy results and is cheap
Method applicable to easily accessible lesions
Modern imaging techniques mainly US & CT applied to
remote sites
Low risk of complications
Can be performed as OP procedure and in patients who
have other co morbidities.
No anaesthesia required
Early diagnosis –helps in diagnostic & therapeutic planning
Cytology-Disadvantages
Small area from the lesion is sampled
Diagnostic difficulty in less cellular lesions
If only necrotic material obtained-diagnosis difficult
Well differentiated cancers may be difficult
Areas of dedifferentiation -can be missed
Needle track haemorrhage/necrosis & reactive
atypia, metaplasia can mislead- esp. in salivary
gland & thyroid
Serious complications –such as major haemorrhage
septicemia, bile peritonitis, a/c. peritonitis,
pneumothorax etc -extremely rare
5. Cassette
Tissues which reach the pathology lab go through a process called
Grossing during which the required area is sliced out into thin
sections of 2x1.5 cm and placed within a plastic container called
the Cassette .Cut pieces of tissue removed during
surgery are put in this for tissue processing
This casette has a lid which can be opened and
the tissue is placed inside and then the lid is closed.
The identity of the patient is written on the
cassette
There are multiple holes in the cassette for
adequate action on the tissue of the various
reagents used during Tissue Processing
6. Paraffin Block
During the processing of tissues, after they have
undergone Dehydration, Clearing and
Impregnation, the tissue is now transferred
into a mould filled with molten wax. This is
called the Paraffin block. The tissue should be
properly oriented while preparing the
[Link] part of the cassette which has the
patient identity number is stuck to this block
of molten wax. The block is quickly cooled
This process of incorporating the tissue into
molten wax is called Embedding. Other agents
like Resin, Gelatin can also be used for
Embedding
7. L Block
Leuckharts Mould Method
This is the traditional blocking
method. The ‘L’ shaped moulds can
be adjusted according to the shape
and size of the tissue to be
embedded. Glycerin may be applied
to the ‘L’ pieces and also to the metal
or glass plate on which the moulds
are arranged before pouring the
molten wax. The tissue is then
embedded. After cooling, the
moulds are removed.
Ice trays, plastic type, are also used to
make tissue block.
8. Block Holder
• This helps to hold the paraffin
block with tissue onto the
microtome which is used for
cutting thin sliced ribbons of
Tissue of 4-6 microns thick
9. Sahli’s Pipette
This is used to draw blood upto 20 mark and
pour it into the Sahli’s hemoglobinometer.
Which is used for determining Hemoglobin
10. RBC Pipette
Made of glass. Has a bulb with a red bead inside.
Red bead helps us to identify it as RBC pipette, also free movement
of the bead means that bulb is dry and the bead helps in mixing
blood with diluting fluid
Uses a) RBC count b) WBC count in severe Leukocytosis(Leukemias)
c) Platelet count
11. WBC Pipette
• Made of glass. Has a bulb with a white bead inside.
• White bead helps us to identify it as WBC pipette, also
free movement of the bead means that bulb is dry and
the bead helps in mixing blood with diluting fluid
• Uses a) WBC count
b) RBC count in severe
anemia
c) Sperm count
d) CSF cell count
[Link] Chamber
The counting chamber contains two ruled areas. These
ruled areas are so small that can not be seen with the
naked eye. Both ruled areas are identical.
Empty space or moat separates the two ruled areas.
There are two raised ridges on either side of the ruled area.
When the cover slip is placed on these elevated ridges, it
acts as a roof covering both ruled areas.
Between the cover glass and ruled areas, there is a tiny
distance of 0.1mm.
MEASUREMENT:- 1 mm x1 mm x 0.1mm = [Link]
USES:- WBC count, RBC count, Platelet count,
Sperm count,CSF count
Counting chamber
13. Wintrobes tube
• This is a glass tube of 11cm length
and 3mm diameter One end is open
while the other is closed
• It is calibrated from 0 to 10 cm and
this is equivalent to 0 to 100 mm
• Used to determine both PCV and
ESR
• From Zero down to top reading is
taken for PCV while zero on top to
down reading is taken for ESR
• Anticoagulant used is double
oxalate
14. Westergren’s tube
• It is glass tube of length
30cms, diameter 2.5mm.
Open at both ends. It is
calibrated from 0 at the top
to 200mm at the bottom
• USE:- To determine the ESR
• Anticoagulant used is
Trisodium citrate
15. Urinometer
Used to determine Specific gravity of urine
This instrument consists of a bulb and a
stem which contains a scale calibrated
in specific gravity reading
This is made to float in a cylinder
containing urine. The depth to which it
sinks in the urine indicates the specific
gravity which is read from the scale
Scale has markings from 1.000 to 1.060
• Normal specific gravity of urine -1.003 to 1.005
• 1.010 is known as fixed specific gravity when the kidney
loses its concentrating or diluting capacity as in end
stage kidney failure
• Specific gravity can be measured using Dipstick
• Substances like albumin, glucose can increase the
specific gravity and hence suitable correction should be
done
• The instrument is calibrated for use at 15.5 degrees
Fahrenheit . In rooms with temperatures above and
below this temperature, suitable corrections are to be
made.
Vacuettes
Blood Bag
Urine-Dipstix
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