restained. FNAC (FINE-NEEDLE ASPIRATION CYTOLOGY) Transcutaneous Aspiration of Palpable Lesions A 20 mL plastic disposable syringe with 21 to 23-gauge
Two liter automatic tissue processor
¾ Simple to use electronic timer and program controls
provide virtually unlimited programing capacity with
¾ Capacity of 160 cassettes in double load or up to 336
¾ Antievaporation lids on all beakers reduce fume levels
¾ Two independent program memories can store up to 4
¾ Vacuum impregnation during processing cycle if
optional vacuum pump utilized.
One liter automatic tissue processor
¾ Economic versatile processing for single or double load
¾ 12 station cycle—with 9 or 10 one liter beakers and 2 or
3 thermostatically controlled wax baths
¾ Automatic agitation ensures efficient impregnation of
¾ Wide range of tissue baskets—large capsule and divided
types in stainless steel and plastic
¾ Standard 24 hours timer plus 24 hours delay.
¾ Exceptional strength and stability with heavy weight
construction, which eliminates vibration and chatter
¾ Unique linear bearing action provides smooth almost
¾ Sections paraffin wax embedded tissue blocks
consistently and accurately from 0.5 to 30
¾ Unique, easy to adjust knife holder accepts wide range
of knives including glass, tungsten carbide tipped, and
¾ Optional lightweight knife holder provides additional
Automatic microtome knife sharpener
¾ Rapid high quality consistent sharpening of microtome
knives, no special training required
¾ Designed to sharpen tungsten carbide tipped as well as
¾ Wide range of knife holders available for C type, D type
¾ Easy to use damper; touch button controls and digital
¾ Unique, hard wearing microsharp iron lapping plate
¾ Easy to adjust facet angle indicator.
Random access stainer for histology
¾ Special built-in fume extraction system, protects
laboratory personnel from xylene fumes
¾ Provides consistent uniform staining of specimens on
¾ Random access provides flexibility to use various
staining routines and eliminates batching
¾ Allows free choice of stain source and type
¾ Running water wash selectable at all positions
¾ Proven, reliable design with a few moving parts
¾ Compact, narrow shape allows wise use of bench space.
Automatic 24 position batch stainer
¾ Simple-to-use electronic timer and program controls
provide virtually unlimited programing capacity with
¾ Immersion times can be programed to the second with
all the versatility of hand staining
¾ Individual immersion times can be reprogrammed to
take account of changing conditions
¾ Two independent program memories can store up
to 4 complete staining routines. Three capacities of
stainless steel slide carrier available—10 slide, 40 slide
¾ Compatible with Autoslip automatic over slipper.
¾ Designed for H and E staining and hematological
¾ Equal or unequal times can be selected.
¾ 224 slides per hour output on continuous loading
¾ Single load operation for 16 or 32 slides horizontally or
¾ 60 minutes timer with additional option for 30 minutes,
¾ Aeration and agitation provided.
¾ Applies mountant and coverslip to standard microscope
slides automatically, presenting the finished slide for
¾ Produces coverslipped slides of consistent high quality
at the rate of up to 160 slides per hour.
¾ Coverslipped slides held in xylene saturated atmosphere
to prevent drying out of specimens prior to coverslipping.
¾ Unique plastic slide clips enable slides to be transferred
direct from Varistain 24 series of automatic stainers.
Cytology is that branch of diagnostic medicine, which deals
with the study of individual cells and/or tissue fragments
spread on laboratory slide and stained appropriately.
Stains used commonly are Papanicolaou’s stain and
May-Grünwald-Giemsa (MGG) stain. For the former,
alcohol fixation is required; and the latter, fixation should
be done by using methanol. For acid-fast bacilli staining,
ethanol fixation is used. Wherever, ethanol fixation is
required, the material obtained and spread on a slide
should be immediately immersed into ethanol. For MGG
staining, the smears are air dried and methanol fixed.
relatively watery, thin and hypocellular fluids need
centrifugation, the sediment so obtained is smeared on to
the slides, stained, and examined.
The advantage of cytological techniques is the rapidity
with which the diagnosis can be provided. However,
this branch has not, and cannot, replace the ultimate in
diagnosis, namely—histopathology.
Cytological study can be done on discharges from the
body (vaginal, nipple, sinus, etc.), scrappings obtained
(from buccal mucosa, or other mucosal surfaces
approachable by employing fiber-optic endoscopes), or by
aspirating from palpable lumps (abscesses, growths, etc.).
In any case, the material obtained is smeared on slides, an
easy and convenient way is to put the material between
two slides and pull them apart or smearing on the slide
by using a coverslip with application of gentle pressure.
All liquids (relatively hypocellular, e.g. urine, body cavity
fluids, etc.) have to be centrifuged, the sediment is used for
smearing. If a bigger chunk of cellular material is obtained,
it can be submitted for histopathological examination
too. Do not forget to add EDTA to containers in which
coagulable fluids are to be collected.
Cytological diagnosis is an important part of cervical
(gynecological) lesions, accessible mucosal lesions and
soft tissue tumors palpable superficially or else approached
under fluoroscopic guidance. Using vacuum to suck and
needle movement in various directions (to dislodge tissue
fragments), sufficient amount of material can be obtained.
The commonly used stains are Pap’s stain and MGG.
Details of Pap’s staining are mentioned below. MGG
staining is similar to that of blood peripheral smear
PAPANICOLAOU METHOD OF STAINING
1. Exfoliated cells degenerate rapidly; therefore, smear
should be prepared and fixed immediately. If there is
to be any delay, the specimen should be fixed in 95%
alcohol and refrigerated until smears can be prepared.
Specimens requiring centrifugation (e.g. urine and
various fluids) are preserved by adding an equal
volume of 50% ethyl alcohol, centrifugation is at 2000
2. Viscid secretions (e.g. vaginal, cervical and prostatic)
should be smeared directly onto clean glass slides and
3. Body fluids and watery exudates (e.g. urine, spinal
fluid, pleural fluid, etc.) will not adhere to the glass
slides unless the slide is first coated with a layer of
Mayer’s egg albumin (one drop per slide).
4. The sediment or centrifuged specimen is smeared
onto glass slides coated with Mayer’s egg albumin.
Any remaining sediment should be processed
as a biopsy specimen for conventional histology
1. Use equal parts of ether and 95% alcohol.
2. Smears should be fixed immediately while still
wet, though partial drying along the edges may be
permitted to prevent the material from becoming
3. Fixation time is 30 minutes to 1 week.
4. If unstained slides are to be mailed, they must be
placed in the fixative for at least 2 hours, then dried
and placed face-to-face before shipment.
1. Harris’s Hematoxylin (modified)
• Ammonium or potassium alum 100 cc
For this purpose, acetic acid should not be added.
• Orange G6, 0.5% solution in 95%
• Phosphotungstic acid 0.015 g
equalled by mixture, prepared in laboratory.
1. After fixation, transfer slides without drying directly
from alcohol—ether solution to 95% alcohol, then
through 80% alcohol, 70% alcohol, and water, to
2. Stain in Harris’s hematoxylin (modified), 8 minutes.
3. Rinse gently in tap water to prevent cells from being
4. Differentiate carefully the nuclear staining in 1%
hydrochloric acid in 70% alcohol; the nuclei should
be clear and sharp in detail; the cytoplasm should be
5. Place in gently running tap water for 5 minutes to
wash out the acid thoroughly and to blue the nuclei.
6. Rinse in distilled water and transfer through 70%
alcohol, 80% alcohol, to 95% alcohol.
7. Stain in OG 6 for 2 minutes.
b. Once in 1% acetic acid in 95% alcohol
c. Once in 1% phoshotungstic acid in 95% alcohol
9. Stain in EA 50 for 3 minutes.
b. 2 changes of absolute alcohol
Nuclei—blue with clear sharp details.
Cytoplasm—varying shades of pink, blue, yellow, green
gray. If necessary, slides may be decolorized in acid alcohol,
washed thoroughly in tap water to remove all acid and
FNAC (FINE-NEEDLE ASPIRATION CYTOLOGY)
Transcutaneous Aspiration of Palpable Lesions
A 20 mL plastic disposable syringe with 21 to 23-gauge
fine needles of variable length, depending upon the
site of tumor, are used for aspiration. The syringe is
fitted with especially designed handle, which permits
a single hand operation during aspiration. The skin is
cleaned with antiseptic solution. No local anesthesia is
required. The tumor mass is fixed with one hand and
with the other hand aspiration is carried out. When
the needle enters the tumor, the plunger of the syringe
is retracted to create a vacuum in the barrel and the
needle is moved to and fro 3 to 4 times. For adequate
sampling, the needle may be moved in three to four
different directions (Fig. 26.1). After completion of
aspiration, the plunger is released before taking out the
needle in order to equalize the pressure. The needle is
disconnected and after filling the syringe with air, it is
reconnected. The content of the needle is expressed on
clean glass slides. Smears are made by applying a gentle
pressure with the flat surface of another glass slide and
allowed to air dry. Alternatively, the flat surface of a
covership can be used to prepare the smears. Smears
are routinely fixed in methanol for MGG staining.
Whenever Pap staining is required for better nuclear
cytochemical stains when fluid aspirated, is discharged
into a clean tube and centrifuged at 1500 rpm. Smears
are made from the deposits when the aspiration fluid
is admixed with blood or frankly hemorrhagic, it may
be collected in a heparinized container. Hematocrit
method or lymphoprep can be used to separate the
tumor cells from RBCs. In case, where cellularity is poor,
the deposit is resuspended in 1 mL of supernatant, spun
in a cytocentrifuge and processed in a similar manner.
Aspiration cytology of intrathoracic masses is usually
performed to diagnose peripheral lung lesions, which
are not accessible by bronchoscope and which do not
desquamate into the bronchial tree. This procedure is
usually carried out around the table under television
fluoroscopy. The skin is cleaned with iodine and spirit and
infiltration with local anesthesia is done up to pleura. The
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