HSBTE 2nd Semester DMLT Unit II. Collection and Preservation of Clinical Specimens for Biochemical Analysis of Body Fluids.

BODY FLUIDS

Cerebrospinal fluid Collection

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 the 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.

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 4 cm to 5 cm.

• 3. Collect the fluid: Once the 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.

• · It may be stored in the refrigerator at a temperature of 2 to 8 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.

Semen/Seminal fluid collection

Seminal fluid is a viscid, neutral, or slightly alkaline, pale-colored fluid containing spermatozoa. It is collected through the ejaculation process.

Purpose to collect seminal fluid:

• 1. Fertility evaluation in males.

• 2. Reproductive system disorder diagnosis.

• 3. Reproductive technology assistance such as IVF (in-vitro fertilization).

The following steps are followed to collect seminal fluid.

• 1. Abstain from sexual activity: Seminal fluid is collected after continuously controlling sex desire for three to five days. This allows a higher concentration of sperm in the seminal fluid.

• 2. Collect the sample: Collect the sample either by masturbation or using a special condom during sexual intercourse to collect ejaculate. Most commonly it is collected in a wide mouth and dry container 50 ml capacity after ejaculation.

Semen/Seminal fluid preservation

It is preferred to examine semen immediately after its collection or within 30 minutes to 1 hour. If storage is required, it should be stored at body temperature (37oC) without using any preservative. Never store in the refrigerator.

• Semen specimens should neither be collected in a condom nor stored in a condom. The condom contains powder and lubricants that affect sperm activity and semen composition.

• Sputum collection: Sputum is the mucus that is colorless, odorless bronchial secretion. It consists of 90% water and 5% soluble components.

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

• Process to collect sputum

  • · Sputum specimen is collected in the early morning hour.

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

  • · Take a deep breath

  • · Coughed up sputum from bronchi.

  • · Place cough carefully in a sterile wide-mouth sterile leak proof glass container with a capacity of 50 to 60 mL.

  • · Avoid mixing sputum with saliva.

  • · Label the container with the name, date of birth, and time of the sample collection.

• Sputum preservation:

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

  • 2. 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.

  • 3. 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 preservation of sputum sample shall also depend upon the specific test to be performed.

Pleural Fluid collection

Pleural is present inside the pleural cavity. The pleura 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 thoracentosis.

The following steps are followed to collect pleural fluid samples:

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 the ribs and into the 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 leakproof 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.

Pericardial Fluid collection: 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 leakproof 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.

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 leakproof 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.

• Synovial Fluid Collection: Synovial fluid is a thick, viscous slippery liquid found in the joints. It allows smooth movement in the joints.

• Purpose of synovial fluid collection: To diagnose various conditions of joints such as arthritis, inflammation or infection, presence of WBC, bacteria, crystals, etc in synovial fluid,

The following steps are followed to collect a sample of synovial fluid

• 1. Pre-procedure preparation: Ask the patient to 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 long thin needles into joints. 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 a bandage at the puncture site to stop bleeding.

• 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 synovial fluid

Transfer the sample into a sterile leakproof 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.

Dr Pramila Singh.