Are you curious about how frozen sections are performed? Many people have heard the term but have no idea about the process. Continue reading to learn more about this fascinating technique.
What Is a Biopsy?
A biopsy describes a small sample of tissue that’s cut from an abnormal appearing mass or tumor. It’s then examined under a microscope by a pathologist. Using the information found, the pathologist provides a diagnosis.
The Role of Frozen Sections in Diagnosing Disease
A frozen section describes one method used for processing biopsied tissue. This is only performed if it will change decisions about the surgical procedure or post-op care.
How Frozen Sections Are Performed
The following describes the steps for performing a frozen section biopsy. Different situations require adjustments in the technique.
Initial Assessment
Ensure that the tissue is fresh so that it will adhere to the slide. Determine whether to perform a smear or touch prep. Also, decide if you will freeze the entire specimen or only part of it.
Fixed Tissue
Place the specimen on a slide intended for fixed tissue. Ensure the specimen has been preserved in formalin.
Freezing the Tissue
The tissue is put in an optimal cutting temperature (OCT) embedding media such as Cryogel or TBS. The optimal tissue sample size is between 3 mm and 5 mm. Specimens that are thinner freeze quicker with fewer crystal artifacts.
A chuck is the tissue holder used to put the specimen on a pre-cooled freezing temperature bar. Prepare the chuck by placing it into a -20 to -15-degree cryostat.
Place the tissue onto the prepared semisolid chuck. Quickly add more media over the tissue until it’s completely covered. Place the chuck back inside the cryostat.
Next, apply a heat sink to the top of the tissue. This takes place at a specified point in the process. The purpose is to speed the freezing and flatten the tissue surface into a flat plane.
You must take care to avoid completely freezing the OCT media.
Cryosectioning
After completing the freezing process, place the chuck close to the cutting blade. Make sure to check that the tissue is fully facing the blade. This ensures an even, full slice of tissue with one full revolution of the machine’s wheel.
Use an anti-roll bar to make sure the tissue doesn’t fold or curl. To polish the tissue, rotate the wheel 10 to 15 times. Make sure that the trim setting is off during this process.
When the machine finishes the cut, a brush catches the curling edge of the tissue section. Next, the brush “jumps” off the block with the curl of tissue. The brush pulls the tissue section so that it’s flat over the stage.
A glass slide can then be gently laid on the tissue section and the tissue then melts onto the slide. Next, prepare the slide using formal alcohol, 95% alcohol, or formalin. Avoid any delays at this stage because it can cause the tissue to dry out.
Slide Staining
Apply the proper reagents and stain to the slide. The choice depends on the type of tissue and test being performed. Once the staining is complete, cover the specimen with a coverslip.
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