Exfoliative Cytology: Preparation of Vaginal & Cervical Smears. Collection and Processing of Specimens for Cytology: Urine, Sputum, CSF (Cerebro Spinal Fluid), and Other Fluids.

Exfoliative Cytopathology/cytology

Cytology is the study of cell structure, functions, and processes that occur in the cell. Cytopathology focuses on the examination of individual cells to diagnose diseases at the cellular level. Exfoliative cytopathology is the study and analysis of cells that have been shed from various surfaces of the body. It is a non-invasive process. Cells are collected by brushing, scraping, or washing the surface of tissues or body cavities. Exfoliative cytopathology deals with the study of cell morphology and the identification of cellular abnormalities.

Uses/Role of Exfoliative Cytopathology

The following are the uses of exfoliative cytopathology

• 1. Cancer screening and diagnosis

• 2. Monitoring disease progress and treatment response

• 3. Diagnosis of infection

• 4. Identification of inflammatory conditions

• 5. Detection of hormonal imbalances

• 6. Research and genetic studies.

Preparation of vaginal & cervical smears

A vaginal smear also called a vaginal swab is commonly used for Exfoliative cytopathology studies. The following steps are followed to prepare a vaginal smear.

• 1. Requirements: Sterile swab, speculum, microscopic slides, and light source.

• 2. Positioning of the Patient: Explain the procedure to the patient regarding the preparation of the vaginal smear. Ask for patient consent. Ask the patient to lie down on the examination table for easy access to the vagina.

• 3. Visual Examination: Put on sterile gloves and hold a sterile swab. Gently open the vaginal wall using a speculum to visualize the vaginal wall and cervix.

• 4. Swab Collection: Insert swab into vagina. Rotate the swab inside the vagina against the vaginal wall to collect vaginal cells and vaginal discharge.

• 5. Slide Preparation: Remove the swab from the vagina and transfer to the sterile microscopic slide. Spread the collected vaginal content on the microscopic slide to form a thin even layer. Allow the slide to dry in air at normal temperature.

• 6. Fixation: Use a proper fixative solution to fix cells on the dry slide. Fixation helps to preserve cell morphology and stop their degradation. Overflow 90% alcohol on cells smear for a few minutes to fix the cells.

• 7. Labelling: Write the patient's name, date, and other required information on the one end of the slide.

Collection and Processing of Urine for Cytology

A urine sample for cytology is collected to examine it under a microscope. A urine sample for cytology is collected to diagnose urinary tract infections, bladder cancer, kidney diseases, and other disorders. The following steps are followed to collect urine samples for cytology

• 1. Patient Preparation: Instruct patient to wash genital part properly using mild soap and water to minimize contamination in urine sample.

• 2. Collection of Urine Sample: Urine sample collection methods depend on the purpose of urine examination. The following are the most commonly used methods to collect urine samples.

  • Mid Stream Urine Sample: Instruct patient to urinate and discard the first part of the urine. Collect midstream urine in a sterile container.

  • 24-hour Urine Collection: Instruct patient to collect all urine excreted over a 24-hour period in a sterile container.

  • Catheterization of Urine Sample: If the patient is unable to collect a midstream sample or an upper urinary tract infection is suspected then a urine sample is collected by using a catheter.

• 3. Sample Preservation: the urine sample should be immediately transported to the medical laboratory for further processing. If it is not possible then it should be refrigerated or preserved by using a preservative.

• 4. Processing for Cytology: The following steps are followed to prepare a slide for cytology.

  • Centrifugation: the urine sample is centrifuged to collect cellular contents at the bottom of the centrifugation tube.

  • Re-Suspension: Cellular content in the bottom of the centrifugation tube is mixed with the saline to make a homogeneous suspension.

  • Slide Preparation: A small volume of the above suspension is placed onto one end of a sterile glass slide. Spread it evenly and allow it to dry in the air. If required, it may be stained by using staining techniques such as Papanicolaou (Pap) and May Grunwald & Giemsa staining (MGG) staining, etc.

• 5. Microscopic Examination: The prepared slide is examined under a microscope. Prepare a report on the basis of abnormal cells observed in the sample.

Collection and Processing of Sputum for Cytology

Sputum is colorless and odorless mucus and other materials expelled from the respiratory tract through coughing. It consists of 90% water and 5% soluble components. Sputum for cytology is examined under a microscope to detect abnormal cells present in the lungs and respiratory tract.

Purpose of Collect Sputum: To diagnose respiratory tract cancer cells, infections caused by bacteria, viruses, fungi, mycobacterium, etc, or other abnormal cells in smokers or lung cancer.

The following steps are followed to collect and process sputum for cytology.

• 1. Patient preparation: Advise patient to rinse mouth properly with water to remove food particles and debris. Otherwise, they will contaminate the sputum sample.

• 2. Sputum sample collection: Sputum collection is preferred in the early morning hour. Instruct patient to

  • § Rinse mouth properly by using potable water to remove food debris and food particles.

  • § Take a deep breath and expel cough forcefully. This helps to collect sputum from the depth of the respiratory tract (Bronchi).

  • § Not to mix sputum with saliva during expulsion of cough.

  • § Place sputum in a sterile wide mouth and leakproof container supplied by the medical laboratory.

  • Write the name, date, time of sample collection, and any other required information on the label of the container.

• 3. Sample Transportation: Assure the container to prevent leakage and contamination of the sample during transportation of the sample. Maintain the integrity of the sample during transportation.

• 4. Sputum Preservation:

  • Refrigeration: Refrigerate the sputum sample immediately after collection in a sterile container. This preserves the integrity of the sample.

  • Freezing: Frozen the sample, if it is to be stored for longer periods. Freeze it at -70 degrees C to avoid repeated freeze-thaw cycles.

  • Fixative: Add fixative, if the sample is to be transported for a longer distance or to be stored for a longer duration. Examples of fixatives are 10% formalin, 70% ethanol, and 2.5% Glutaraldehyde. Types of fixatives of sputum samples shall also depend upon the specific test to be performed.

• 5. Sample Processing: The following steps are followed to process and collect samples for cytological examination

  • Inspect the quality and volume of the sputum sample.

  • If required, concentrate the cells present in the sputum sample by centrifugation.

  • Prepare the smear of the sputum sample onto a sterile glass slide and label it properly.

• 6. Staining and Fixation: Stain smears using the staining technique to enhance the visibility of the cells under a microscope. Papanicolaou (Pap) stains,  Romanowsky stains, or Wright’s stains are commonly used.

• 7. Microscopic Examination: A stained and dry slide is examined under the microscope. Detect the abnormal cells in the smear and report it to healthcare provider.

Collection and processing Cerebro Spinal Fluid:

The process to collect CSF is called lumbar puncture or spinal tap. CSF specimens should be collected by physicians or trained medical laboratory technicians or nurses. CSF is a clear colorless liquid that surrounds the brain and spinal cord.

Purposes:

• 1. Diagnostic purposes: To diagnose

  • Meningitis: Inflammation of membranes that surround the brain and spinal cord,

  • Encephalitis: Inflammation of the brain.

  • Subarachnoid hemorrhage,

  • Intracranial hemorrhage,

  • Intracranial lesion,

  • Multiple sclerosis,

  • Brain tumors: abnormal growth of tissue brain,

  • Hydrocephalus: Accumulation of CSF in brain

• 2. Therapeutic purposes:

  • Relief from intracranial pressure.

  • Administration of serum such as Anti-meningococcus serum, Anti-tetanus serum

  • Administration of Hypertonic solution of magnesium sulfate, and sodium chloride.

  • Administration of lipiodol,

  • Administration of anesthesia.

  • Ventriculography.

Collection of CSF Sample: The following steps are followed to collect the sample of cerebrospinal fluid.

• 1. Pre-procedure preparation: Instruct the patient to lie on his/her side with the knee tucked up to the chest. Or to sit on the edge of the bed or examination table.

• 2. Insert the needle: Select the site by using ultrasound guidance or chest palpation. Sanitize the area of the skin to be punctured. Apply local anesthetic. Insert a thin hollow needle in the space between the 4th and 5th lumbar vertebrae up to A depth of 4cm to 5cm.

• 3. Collect the fluid: Once a needle is in place, connect the needle with a syringe or vacuum to collect cerebrospinal fluid.

• 4. Remove the needle: Remove the needle after collecting a sufficient amount of CSF sample. Apply bandage at the puncture site.

• 5. Post-procedure care: Instruct patient to be in clinic or hospital under observation for a few hours to check for any sign of complication.

• 6. Labeling: Label the sample container with the name of the patient, date of birth, time of collection, and date of collection.

Cerebrospinal fluid preservation:

• · CSF specimens should be examined immediately or within an hour after their collection.

• · It should not be stored in a refrigerator especially if it is collected for bacterial culture purposes. of

• · It may be stored in the refrigerator at a temperature of 2 to 80 degrees C for 2 to 3 hours, if the CSF specimen is collected for biochemical examination.

• · The CSF collection process is difficult and painful. Thus it should be examined carefully and economically.

• · CSF may contain virulent organisms, thus it should be handled with care.

Collection and processing of Pleural Fluid: Pleural is present inside the pleural cavity. Theleura is a double-layered membrane that surrounds the lungs. The inner layer remains in contact with the lungs and the outer layer with the chest cavity. The space between these two layers is called the pleural cavity. the pleural cavity is filled with pleural fluid.

Purpose to collect pleural fluid: To diagnose pleural effusion. Abnormal accumulation of the pleural fluid in pleural space is called pleural effusion.

Process to collect pleural fluid: The process to collect the pleural fluid sample is called thoracentesis. The following steps are followed to collect the pleural fluid sample:

• 1. Pre-procedure preparation: Instruct the patient to sit upright and lean forward on the edge of the bed or table. Instruct patient to take shallow breathing and hold breath during needle insertion.

• 2. Insert the needle: Select the site by using ultrasound guidance or chest palpation. Sanitize the area of the skin to be punctured. Apply local anesthetic. Insert A thin hollow needle between ribs and into pleural space.

• 3. Collect the fluid: Once the needle is in place, connect the needle with A syringe or vacuum to collect pleural fluid. Ask the patient to change position to maximize the pleural fluid collection.

• 4. Remove the needle: Remove the needle after collecting a sufficient amount of pleural fluid sample. Apply bandage at the puncture site.

• 5. Post-procedure care: Instruct patient to be in clinic or hospital under observation for a few hours to check for any sign of complication.

• 6. Labeling: Label the sample container with the name of the patient, date of birth, time of collection, and date of collection.

Pleural fluid sample preservation: Transfer the sample into a sterile leak-proof container. Refrigerate it immediately at 4 degrees C. Do not allow to freeze it. Ensure their transportation to the laboratory in an ice bag to maintain a temperature of 4 degrees C. Do not freeze the sample. Assure to analyze the sample within 24 hours of the collection.

Collection and Processing of Pericardial Fluid

Pericardial fluid is present in the pericardial cavity. The pericardium membrane surrounds the heart. It is made of double layers. The space between these layers is called the pericardium cavity. Pericardial fluid is a clear, yellowish fluid present in the pericardial space to protect and lubricate the heart. The process of collecting pericardial fluid is called pericardiocentesis.

Purpose to collect pericardial fluid: Pericardial fluid is collected for both diagnostic and therapeutic purposes.

1. Diagnostic purposes:

• Pericarditis: Inflammation of pericardium

• Pericardial effusion: Accumulation of excess pericardial fluid of in pericardium.

Cancer: Detection of cancer spread up to the pericardium.

2. Therapeutic purposes:

• Drainage of excess pericardial fluid

• Administration of medication into pericardial space.

Process to collect pericardial fluid: The following steps are followed to collect pericardial fluid.

• 1. Pre-procedure preparation: Ask the patient to lie down on his/her back. Sanitize the area of the skin to be punctured. Apply local anesthetic. Select the site by using ultrasound guidance or chest palpation. Sanitize the area of the skin to be punctured.

• 2. Insert the needle: Apply local anesthetic. Insert a thin hollow needle between the ribs and into the pericardium space. An ultrasound machine or other imaging machine is used to ensure the correct position of the needle during and after insertion.

• 3. Collect the fluid: Use a syringe to withdraw pericardial fluid. The amount of pericardial fluid to be collected depends upon the reason for sample collection.

• 4. Remove the needle: Remove the needle after collecting a sufficient amount of pericardial fluid sample. Apply bandage at the puncture site.

• 5. Post-procedure care: Instruct patient to be in clinic or hospital under observation for a few hours to check for any sign of complication.

• 6. Labeling: Label the sample container with the name of the patient, date of birth, time of collection, and date of collection.

Preservation of pericardial fluid: Transfer the sample into a sterile leak-proof container. Refrigerate it immediately at 4 degrees C. Do not allow to freeze it. Ensure their transportation to the laboratory in an ice bag to maintain a temperature of 4 degrees C. Do not freeze the sample. Assure to analyze the sample within 24 hours of the collection.

Collection and Processing of Peritoneal Fluid

The peritoneum is a membrane to surrounds the abdominal cavity. Peritoneal fluid is present in the peritoneum. The process of collecting peritoneal fluid is called paracentesis. The following steps are followed to collect peritoneal fluid.

• 1. Pre-procedure preparation: Ask the patient to lie down on his/her back or sit depending upon comfort level. Sanitize the area of the skin to be punctured. Apply local anesthetic. Select the site by using ultrasound guidance or chest palpation. Sanitize the area of the skin to be punctured.

• 2. Insert the needle: Apply local anesthetic. Insert a long thin needle into the peritoneal cavity. An ultrasound machine or other imaging machine is used to ensure the correct position of the needle during and after insertion.

• 3. Collect the fluid: Use a sterile syringe to withdraw peritoneal fluid.

• 4. Remove the needle: Remove the needle after collecting a sufficient amount of peritoneal fluid sample. Apply bandage at the puncture site.

• 5. Post-procedure care: Instruct patient to be in clinic or hospital under observation for a few hours to check for any sign of complication.

• 6. Labeling: Label the sample container with the name of the patient, date of birth, time of collection, and date of collection.

Preservation of peritoneal fluid: Transfer the sample into a sterile leak-proof container. Refrigerate it immediately at 4 degrees C. Do not allow to freeze it. Ensure their transportation to the laboratory in an ice bag to maintain a temperature of 4 degrees C. Do not freeze the sample. Assure to analyze the sample within 24 hours of the collection.