HISTOPATHOLOGY

Pathology seems to be familiar, for we hear mostly of it in association with diseases, but for laymen, less
is known about what histopathology is. Divided into two major branches, pathology has gross pathology
and microscopic pathology. Microscopic pathology consists of two branches: clinical pathology, which
involves clinical chemistry, clinical microscopy, immunology, serology, bacteriology, and hematology,
among others, and anatomical pathology, which solely focuses on histopathology. Histopathology
involves microscopically observing and examining the changes that the tissue samples present, which
are indications of a disease. In the laboratory, medical technologists work in this department to assist the
pathologist by performing the different tissue processes and histopathologic techniques, from fixation to
mounting and labeling. In some instances, as is the case here in the Philippines, pathologists allow
medical technologists to perform gross examinations. Histopathology underscores a remarkable and
efficient detection of disease since disease processes affect tissues excessively and manifest in their
minute appearance. An alteration of tissue from its normal state can be used as the basis for an
accurate diagnosis of a health condition that will allow the physicians to administer effective treatment
for the patients. Hence, histopathology is a crucial component of medicine and healthcare services,
owing to its concentration on the study of diseases of tissues.

DEHYDRATION
Dehydration is the next step following fixation for tissues that do not necessitate being decalcified, and it
is the subsequent step for specimens like bones, teeth, or blood vessels with calcium deposition due to
atherosclerosis, which undergo decalcification. It is a process of removing intercellular and extracellular
water from the tissue that involves slow substitution with an organic solvent in preparation for
impregnation. This step is performed by utilizing dehydrating agents in graded series for short periods of
time to reduce the possibility of shrinkage and extraction of cell components since abrupt osmotic
changes brought about by the replacement of water can generate distortions, which must be prevented.
To better remember and understand the importance of this process, it is considerable to think that
AFTER being fixed, one must ensure to face the REMOVAL of any EXCESS water of emotions inside
before taking on another journey. Likewise, the tissue, after being fixed, must proceed to the total
removal of excess water to ensure its readiness for the subsequent process.

Choosing a dehydrating agent is a vital consideration for a good-quality tissue sample. A dehydrating
agent that dehydrates rapidly even fatty tissues, does not evaporate swiftly, does not harden tissues,
does not remove stains, is not toxic, and is not a fire hazard is definitely deemed an ideal dehydrating
solution. Alcohol, especially ethanol, is the most common dehydrating agent that can be utilized for
effective water removal, while isopropanol is a close substitute. Other dehydrating agents such as
acetone, dioxane, cellosolve, triethyl phosphate, and tetrahydrofuran can also be used.

CLEARING
Since dehydration has already rendered the tissue water-free, it is necessary to further process the
tissue to make it ready for infiltration. A dehydrating agent such as ethanol is immiscible with paraffin
wax, which is routinely used for infiltration; hence, an intermediate solvent that is miscible with both is
imperative. This process is named as such due to its capacity to impart transparency or optical clarity to
the tissue samples, enabling a more detectable appearance necessary for examining the tissue under
the microscope. The alteration in appearance indicates the completeness of the clearing process, which
gives a high refractive index. In addition, this step eliminates a significant quantity of fat in the tissue that
can impede the wax from infiltrating it once it is not effectively removed. In choosing an effective clearing
agent, it is crucial to consider the type of tissues to be processed and the type of processing to be
undertaken. This step in tissue processing can be efficiently grasped if it is compared to a real-life
phenomenon that is palpable in situations where, after excess emotions are removed, clearing one’s
mind is essential. When every unnecessary thing is already REMOVED, one seeks clarity of mind,
which is achieved by experiencing transparency within oneself. Tissue samples, as part of human
beings, must also be cleared after the removal of excess water. For an effective CLARITY and
TRANSPARENCY of tissues renders a good infiltration.
Opting for a good clearing agent requires that it be miscible with alcohol and paraffin wax and can make
tissue apparent. It must not produce excessive shrinkage, hardening, or damage to tissue. Also, it
should not dissolve aniline dyes and should not evaporate quickly in a water bath.

LABELING
Prior to actual viewing of tissue samples under the microscope and even before the tissue processing
starts, labeling is an essential step to perform with utmost correctness to ensure accuracy of results
given due to the rightful patient. Name of the patient, the sex and other personal information must be
checked to confirm the identity of the patient and type of specimen and the date the sample is submitted
as well as the type of test indicated in the requisition form must also be validated.