Collection of samples

 Female genital tract, Respiratory tract, Urinary system, GIT, Skin and
oral cavity, breast nipple discharge
 Body fluid
 Fine needle aspiration cytology, USG guided FNAC
Collection of sample from female genital tract
• Proper specimen collection and submission is very important.
• At least one half to two thirds of false negatives are the result of
patient conditions present at the time of sample collection and
submission and the skill and knowledge of the individual who obtains
the specimen.
• Thus sample collection is very important and health personnel
responsible should be trained enough.
• The laboratory provides feedback on sample adequacy via individual
reports, and may elect to provide summary information regarding
patient sampling to its clients.
• Patient Preparation
 To optimize collection conditions, a woman should:42
 Schedule an appointment approximately two weeks (10-18 days) after
the first day of her last menstrual period.
 Not douche 48 hours prior to the test.
 Not use tampons, birth control foams, jellies or other vaginal creams
or vaginal medications for 48 hours prior to the test.
 Refrain from intercourse 48 hours prior to the test.
• Test Requisition
• Under the supervision and guidance of the physician, a laboratory requisition
must be legibly and accurately filled out before obtaining the cellular sample.
• Patient’s name
• Age
• Menstrual status (LMP, hysterectomy, pregnant, postpartum, hormone therapy.)
• Previous abnormal cervical cytology result, previous treatment, biopsy or surgical
procedure.
• Patient’s risk status for developing cervical cancer, e.g. “high risk”.
• Source of specimen, e.g. cervical, vaginal.
• Hormone/contraceptive use.
• Relevant clinical findings (abnormal bleeding, grossly visible lesion, etc.)
• Labeling the Sample
• The glass slide or specimen vial must be labeled with a unique
identifier, usually the patient’s first and last names, at the time of the
collection of the cellular sample.
• For glass slides, the required information is written in solvent
resistant pen or pencil on the frosted end of the slide. For liquid
based samples, the required information must be affixed to the vial.
• Visualization of the Cervix for Collection of an Adequate Sample
• Collection of a cervical cytology specimen is usually performed with the patient in the lithotomy
position.
• A sterile, or single-use bivalve speculum of appropriate size is inserted into the vagina without
lubrication.
• The position of the speculum should allow for complete visualization of the os and ectocervix.
• The transformation zone is the site of origin for most cervical neoplasia and should be the focus
of cytology specimen collection.
• The transformation zone may be easily visualized or may be high in the endocervical canal.
• Location varies not only from patient to patient, but in an individual over time. Factors producing
variation include changes in vaginal pH, hormonal changes including pregnancy, childbirth, and
menopausal status, and hormonal therapy.
• If a patient has had a hysterectomy, a vaginal sample is sufficient, with particular attention to
sampling the vaginal cuff.
• Collection Devices
• There are a variety of collection devices available for sampling the endocervix,
transformation zone and ectocervix.
• They include endocervical brushes, wooden and plastic spatulas, and plastic
“broom-type” samplers.
• Plastic spatulas are preferred over wooden since the wooden spatulas retain
cellular material.
• The use of a cotton-tipped swab is NOT recommended, even if the swab is
moistened.
• Cells adhere to the cotton and do not transfer well to the glass slide, which
results in an incomplete specimen. Analysis of different sampling methods has
shown that overall, the cytobrush and spatula together provide the best
specimen for cervical
• Cell Fixation for Conventional Cervical Cytology
• Immediate fixation of the cellular sample, within seconds of specimen collection, is necessary to
prevent air-drying.
• Air-drying obscures cellular detail and compromises specimen evaluation. Immersing the slide in
alcohol or spraying with fixative can prevent air-drying artifact.
• If the specimen is immersed in alcohol, it may remain in the alcohol for transport to the
laboratory.
• Alternatively, the specimen can be immersed in alcohol for 20-30 minutes, removed and allowed
to air dry, then placed in a container/mailer for transport to the laboratory.
• The immersion technique requires use of a separate container for each specimen and changing
or filtering the alcohol between specimens.
• If a specimen is spray fixed, only quality controlled cytology fixatives should be used.
• Hair spray should NOT be used.
• Generally, spray fixatives should be 6-10 inches (15-25 cm) from the glass slide when applied.
Respiratory Cytology
• Major role:
– Diagnosis of malignant neoplasms involving lung both primary and
metastatic
• Minor role:
– Opportunistic infection
– Specific inflammatory process
– Benign neoplasms, some
Respiratory tract
1. Sputum:
This mucoid product of the respiratory tract is produced as a component of the protective
mucociliary clearance mechanism to filter foreign particles and protect the underlying
mucosa.
Examination of sputum has the advantages of being cheap, quick, noninvasive, and easily
repeatable.
Early morning sputum, collected for three consecutive days when the patient wakes in the
morning, gives the best results.
The patient should be instructed to clear the throat, rinse the mouth, and cough deeply
using the diaphragm.
For induction, aerosol of heated hypertonic saline (10–15%) with 20% polypropylene
glycol is inhaled.
The specimen can be sent to the laboratory unfixed (if no significant delay is expected) or
in an appropriate fixative or preservative such as Saccomanno solution.
Advantages of sputum:
• Noninvasive
• Reflect constituents from many regions of lung
• Useful for centrally located malignancies (Squamous cell/Small cell
CA)
• High diagnostic yields: induced sputum, 3-5 samples continuously
examined
• Chronic inflammations: Asthma, COPD
• Respiratory infections
Disadvantages of sputum:
• Alveolar macrophages: lower respiratory tract elements
• Localized lung lesion, peripheral lesion
• Adenocarcinoma, metastatic lesion, lymphoma
• Benign tumor
2. Bronchial Brushings:
A brush can be inserted into the bronchoscope and abnormal areas scraped.
The brush is then retrieved, and the material spread directly onto a glass
slide either along the length of the slide or in small circular area centrally.
Alternatively the brush can be rinsed in fluid to produce a liquid sample for
LBC.
Thirdly, the slides can be made and the brush stored in case required later
for agitation and centrifugation and cytospin.
The slide can be fixed, and Papanicolaou stained or air-dried for MGG
preparations. This procedure is reported to have a specificity and sensitivity
of around 90 %.
3. Bronchial Washings :
Aspirates of bronchial secretions and bronchial washes with normal saline
are collected, usually following the brush.
The material should be submitted to the laboratory for processing as quickly
as possible to avoid any deterioration and loss of cytologic detail.
This differs from the brush procedure and involves introducing saline into the
airway(s) under investigation and then collecting or trapping the fluid drawn
off subsequently.
As with other fluid samples, the material can either be handled further in the
routine LBC manner or by centrifugation or cyto-spinning and slide
preparation or production of a cell block as described previously.
4. Bronchoalveolar Lavage:
• BAL involves the infusion and reaspiration of a sterile saline solution in distal segments of the lung via a
fiberoptic bronchoscope.
• This procedure, originally introduced for the treatment of asthma and alveolar proteinosis, is the method of
choice for investigating diffuse nonneoplastic disorders, including a range of opportunistic and other
infections, and also for peripherally situated tumors, especially bronchioloalveolar carcinoma and
metastases.
• It has become established as a tool for managing lung transplants and has a potential role in the diagnosis
and monitoring a range of interstitial lung diseases including idiopathic pulmonary fibrosis and sarcoidosis,
primarily relating differential cell counts to certain diseases and absolute counts to response to treatment.
• The cell preservation is said to be better than routine washings and sputum samples, though it is
recommended that a sample with an epithelial cell count above 5 % should be considered potentially
inadequate.
• The samples are processed in the same way as bronchial washings. The cellular composition and relative
numbers of various cells are used a means to predict the underlying condition in a range of ailments,
especially those causing interstitial lung disease
5. Transbronchial Fine Needle Aspiration (TBNA):
• This technique is relatively new and is particularly useful for nodal
assessment in the paratracheal and mediastinal regions where
lymphadenopathy is frequently due to granulomatous in flammation,
lymphoma, and metastases, among other causes.
• Complications are rare but include pneumothorax and hemorrhage.
• As a rule, a 19–22-gauge needle is safe and well tolerated.
• Direct smears are usually made with air-dried material being stained
with MGG, while fi xed material is usually stained with Papanicolaou.
6. Endoscopic Ultrasound-Guided FNAs :
• The introduction of the endoscopic ultrasound has meant that
peribronchial lesions can be visualized ultrasonographically during the
bronchoscopy and needle inserted to allow more accurate sampling.
• Small lymph nodes and masses that could not be accurately punctured
previously can now be sampled without too much dif fi culty.
• The material is spread on to slides directly or introduced into liquid
medium.
• The solid clotted blood (the “worm”) can be embedded in paraffin and
processed for histology.
• This frequently has signi fi cant fragments of diagnostic tissue and which
can be used for further studies such as immunohistochemistry or
molecular pathology, such as EGFR mutational status.
Urinary system