Cytotechnology and Diagnostic Cytology Lecture Notes PDF

Cytopreparatory Techniques 231 MLT Histology  Cytology Specimen Collection Proper specimen collections Specimen integrity- maintained by proper preservative Sample identification Patient identification- clearly labeled on specimen container and on slide  Specimen Collec...

Cytopreparatory Techniques 231 MLT Histology  Cytology Specimen Collection Proper specimen collections Specimen integrity- maintained by proper preservative Sample identification Patient identification- clearly labeled on specimen container and on slide  Specimen Collections - General Information All cytology specimens received by the laboratory must be accompanied by a completed cytology requisition The following information must be included: Patient's full name Physician(s) name Saudi ID number Date of specimen collection Patient's date of birth Source of specimen (anatomic site) Brief clinical history (CLIA requirement)  Types of cytological specimens 1- Body Fluid (Pleural, Peritoneal, Pericardial, and Synovial) Collect 10-200 ml specimen in a leak-proof, screw-top wide-mouth container Optimum sample size is 100 ml or larger A cell block will be prepared if adequate specimen is obtained  Cervical smears -2 The most acceptable method of obtaining a satisfactory sample is by use of spatula The optimum sample should reach the transformation zone and the squamocolumnar junction of cervix  Gynecological Specimen Collection Pap Smears Collection Smearing  3- Respiratory tract Sputum should be collected on 3 consecutive days Early morning expectorate is preferred before food is eaten or toothpaste used to eliminate contamination The sputum should be from deep cough, not salivary spit, and collected into a sterile container  3- Respiratory tract Bronchoalveolar lavage (BAL), this technique involve introducing approximately 100 ml of sterile saline via bronchoscope BAL technique is used for – Investigation of interstitial lung disease – Detection of pneumocystis carinii pneumonia – Malignant disease Bronchial brushings can also be obtained using a small nylon brush via a bronchoscope  4- Gastrointestinal (GI) tract GI tract specimen obtained by indirect scrapes using endoscopic brushings and ultrasound-guided endoscopic FNA Specimens collected from: – Oral – laryngeal lesions – Esophagus – Gastric brush samples – Rectal and colon  Urinary cytology -5 A full voided specimen of urine is the most appropriate for cytological investigation It should NOT be the first morning sample and NOT a 24-hour collection since cells deteriorate quickly in urine Urine samples should be dispatched without delay to the laboratory for processing Brush samples may be received from bladder, ureteric lesions, and kidney aspirates  Fine-needle aspiration FNA is important and rapidly expanding area of cytodiagnosis Essentially needle aspiration is either of palpable or non- palpable lesions Most common sites for palpable lesions are breast, thyroid, soft tissue and lymph nodes Non palpable lesions need to be identified by some imaging device, such as ultrasound, MRI before FNA introduced to non-palpable lesions  Miscellaneous specimen types Skin scrapes may be taken in cases of basal cell carcinoma Nipple secretions may be collected directly onto glass slides and fixed immediately  Cytological fixatives 1-Alcohol fixation: Most laboratories now used 95% ethyl alcohol for most routine preparations that require fixation Industrial methylated spirit is common substitute for ethyl alcohol  Cytological fixatives 2- Coating fixatives are widely used for cervical smears – Cervical smear is covered in waxy coating to protect the samples from damage Other fixatives: – Composed of polyethylene glycol in alcohol base – Applied using dropper bottle or spray form  Special purpose of fixatives 1- 10% neutral formal saline suitable for fixation of serous clots ad cell blocks 2- Formal vapour is preferred method for lipids 3- Saccomanno’s fixative is composed of alcohol and carbox – Used for preparation of sputum 4- Eposti’s fixative is composed of methanol, D. water and glacial acetic acid – Used for urine equal volume of fixative added to urine. It is 5- Glutaraldehyde fixative used for transmitted electron microscope 6- Air-dried preparation are required for Romanowsky’s stain. 7- Methanol is used for fixing smears that have been air dried  Specimen preparation Direct spread smears: Type of specimens- gynecological, cervix, vagina, and vulva specimens Direct spread smear are collected by wooden or plastic spatula All specimens should be evenly and thinly spread Specimens should be fixed immediately in 95% alcohol  Fine needle aspirates Syringe should be detached from the aspirator and air drawn The material is quite abundant It should be evenly distributed over two slides by placing another slide on top of the one containing the material Separating them quickly by sliding them in opposite direction The smears should be immediately fixed in 95% alcohol before any drying occurs. FNA  CYTOCENTRIFUGATION Urine, serous effusions, and watery lavages required concentration of the cells prior to transfer to glass slides the the specimens should be agitated gently After centrifugation, the cells should be at the bottom of the centrifuge tube The supernatant should be poured off Using a pipette, remove the resulting cell suspension carefully, place in slide and spread material rapidly  Cell blocks Cell block is method of preparing cytology material and process as histology section It can be prepared from fluid specimens of all types but particularly from effusions, endometrial aspirates, brush samples, and FNA Liquefaction is necessary before proceeding The most used method employs 2% agar in 100 ml of distilled water  Cytology staining The universal stain for cytological preparations is the Papanicolaou stain commonly called Pap stain Harris’s haematoxylin is used as a nuclear stain Combination of OG-6 (orange gelb) and EA 50 give the subtle range of green, blue and pink hues to the cell cytoplasm  Romanowsky stains Common use in blood and bone marrow films, and to detect parasites such as malaria Romanowsky stains consist of methylene blue/ azur B and eosin Dissolved in acetone-free methanol Stains that are related to or derived from the Romanowsky-type stains include: Jenner stain Giemsa stain May Grunwald Giemsa stain Leishman stain  Some useful special stains applied to cytological specimens PAS/ diastase Mucicarmine Oil red O Sudan black Masson-fontana Grocott’s methamine silver Immunocytochemistry Best wishes in your midterm exam Why I don’t study earlie