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Vaginal cytology is a microscopic examination of cells from the
vagina In mammals, the vagina is the elastic, muscular part of the female genital tract. In humans, it extends from the vestibule to the cervix. The outer vaginal opening is normally partly covered by a thin layer of mucosal tissue called the hymen ...
l
epithelium Epithelium or epithelial tissue is one of the four basic types of animal tissue, along with connective tissue, muscle tissue and nervous tissue. It is a thin, continuous, protective layer of compactly packed cells with a little intercellul ...
. In veterinary medicine, it helps differentiate the stages of the mammalian
estrous cycle The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous p ...
because the
vaginal epithelium The vaginal epithelium is the inner lining of the vagina consisting of multiple layers of (squamous) cells. The basal membrane provides the support for the first layer of the epithelium-the basal layer. The intermediate layers lie upon the basal ...
changes in response to
sex hormone Sex hormones, also known as sex steroids, gonadocorticoids and gonadal steroids, are steroid hormones that interact with vertebrate steroid hormone receptors. The sex hormones include the androgens, estrogens, and progestogens. Their effects ar ...
levels; practically, it is used to distinguish when a female canine is at a particular point in the estrous cycle. In a normal vaginal smear, lactational cells, navicular cells, endocervical cells, endometrial cells, trophoblastic cells, and
leucocytes White blood cells, also called leukocytes or leucocytes, are the cells of the immune system that are involved in protecting the body against both infectious disease and foreign invaders. All white blood cells are produced and derived from mult ...
may be present. Vaginal cytology allows a clinician to: * Determine whether a canine is actually
in heat The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous p ...
. * Aid in determining the correct time to begin performing more expensive serum
progesterone Progesterone (P4) is an endogenous steroid and progestogen sex hormone involved in the menstrual cycle, pregnancy, and embryogenesis of humans and other species. It belongs to a group of steroid hormones called the progestogens and is the m ...
and
luteinizing hormone Luteinizing hormone (LH, also known as luteinising hormone, lutropin and sometimes lutrophin) is a hormone produced by gonadotropic cells in the anterior pituitary gland. The production of LH is regulated by gonadotropin-releasing hormone (GnRH) ...
(LH) assays for precise
ovulation Ovulation is the release of eggs from the ovaries. In women, this event occurs when the ovarian follicles rupture and release the secondary oocyte ovarian cells. After ovulation, during the luteal phase, the egg will be available to be fertilized ...
timing. * Determine if it is too late in the estrous cycle to perform artificial insemination in dogs unable or unwilling to breed naturally. * Determine if a bitch is under the influence of estrogen. * Predict the correct day to perform an elective
cesarean section Caesarean section, also known as C-section or caesarean delivery, is the surgical procedure by which one or more babies are delivered through an incision in the mother's abdomen, often performed because vaginal delivery would put the baby or mo ...
. The equipment needed for vaginal cytology includes a
vaginal speculum A speculum (Latin for 'mirror'; plural specula or speculums) is a medical tool for investigating body orifices, with a form dependent on the orifice for which it is designed. In old texts, the speculum may also be referred to as a diopter or dio ...
, cotton-tipped applicators, frosted microscope slides, commercial
Romanowsky stain Romanowsky staining, also known as Romanowsky–Giemsa staining, is a prototypical staining technique that was the forerunner of several distinct but similar stains widely used in hematology (the study of blood) and cytopathology (the study of di ...
, and
light microscope The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microsco ...
.


Types of cells

*
Basal cells The ''stratum basale'' (basal layer, sometimes referred to as ''stratum germinativum'') is the deepest layer of the five layers of the epidermis, the external covering of skin in mammals. The ''stratum basale'' is a single layer of columnar o ...
- give rise to all
epithelium Epithelium or epithelial tissue is one of the four basic types of animal tissue, along with connective tissue, muscle tissue and nervous tissue. It is a thin, continuous, protective layer of compactly packed cells with a little intercellul ...
in a vaginal smear. They are small cells with round nuclei, and a high nucleus-to-
cytoplasm In cell biology, the cytoplasm is all of the material within a eukaryotic cell, enclosed by the cell membrane, except for the cell nucleus. The material inside the nucleus and contained within the nuclear membrane is termed the nucleoplasm. The ...
(N:C) ratio. They are rarely observed.Cowell, R., & Valenciano, A. (2019). Cowell and Tyler’s Diagnostic Cytology and Hematology of the Dog and Cat. (5th ed.). St Louis, MO: Elsevier. * Parabasal cells - the smallest epithelial cells seen on a typical vaginal smear, and are derived from the
basal layer The ''stratum basale'' (basal layer, sometimes referred to as ''stratum germinativum'') is the deepest layer of the five layers of the epidermis, the external covering of skin in mammals. The ''stratum basale'' is a single layer of columnar or ...
of squamous epithelial cells. They are round or nearly round and have a high nucleus-to-
cytoplasm In cell biology, the cytoplasm is all of the material within a eukaryotic cell, enclosed by the cell membrane, except for the cell nucleus. The material inside the nucleus and contained within the nuclear membrane is termed the nucleoplasm. The ...
(N:C) ratio. Cytoplasm is dense and
basophilic Basophilic is a technical term used by pathologists. It describes the appearance of cells, tissues and cellular structures as seen through the microscope after a histological section has been stained with a basic dye. The most common such dye is ...
. The nucleus has finely granular
chromatin Chromatin is a complex of DNA and protein found in eukaryotic cells. The primary function is to package long DNA molecules into more compact, denser structures. This prevents the strands from becoming tangled and also plays important roles in r ...
and is vesiculated.Brown, M., & Tighe, M. (2015). Mosby’s Comprehensive Review for Veterinary Technicians. (4th ed.). St Louis, MO: Elsevier * Intermediate cells - two to three times larger than parabasal cells, but their nuclei are similar in size to parabasal cell nuclei. Small intermediate cells are round or almost round with a large prominent nucleus. Large intermediate cells, also called superficial intermediate cells or transitional intermediate cells, have a polygonal shape, and a small N:C ratio. * Superficial cells (dead cells) - the largest cells seen on a vaginal cytology and are derived from the outermost layer of non-
keratinized Keratin () is one of a family of structural fibrous proteins also known as ''scleroproteins''. Alpha-keratin (α-keratin) is a type of keratin found in vertebrates. It is the key structural material making up scales, hair, nails, feathers, horn ...
epithelial cells. They are polygonal in shape, flat, and often have the appearance of being rolled up. The nuclei are absent or pyknotic, meaning small and dark. The cytoplasm may develop small
vacuoles A vacuole () is a membrane-bound organelle which is present in plant and fungal cells and some protist, animal, and bacterial cells. Vacuoles are essentially enclosed compartments which are filled with water containing inorganic and organic mo ...
as the cells age.
Cornification Keratin () is one of a family of structural fibrous proteins also known as ''scleroproteins''. Alpha-keratin (α-keratin) is a type of keratin found in vertebrates. It is the key structural material making up scales, hair, nails, feathers, hor ...
is the degeneration process into these large, flat, anucleated cells. Superficial cells without nuclei are considered fully cornified. *Metestrum cells - vaginal epithelial cells containing
neutrophils Neutrophils (also known as neutrocytes or heterophils) are the most abundant type of granulocytes and make up 40% to 70% of all white blood cells in humans. They form an essential part of the innate immune system, with their functions varying in ...
in their cytoplasm. Although their name implies they are seen during metestrus, these cells are not specific for any stage of estrous. *Foam cells - sometimes seen during
anestrus The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous ...
, and are simply nondescript cells with large vacuoles.


Observable cells in each stage of estrous cycle


Proestrus

* Early- to mid-
proestrus The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous p ...
- During this stage, vaginal cytology will show a gradual shift from parabasal cells, to intermediate cells, to superficial cells. Also present are
neutrophils Neutrophils (also known as neutrocytes or heterophils) are the most abundant type of granulocytes and make up 40% to 70% of all white blood cells in humans. They form an essential part of the innate immune system, with their functions varying in ...
and
red blood cells Red blood cells (RBCs), also referred to as red cells, red blood corpuscles (in humans or other animals not having nucleus in red blood cells), haematids, erythroid cells or erythrocytes (from Greek language, Greek ''erythros'' for "red" and ''k ...
in large numbers. The red blood cells are due to the proliferation of
vaginal epithelium The vaginal epithelium is the inner lining of the vagina consisting of multiple layers of (squamous) cells. The basal membrane provides the support for the first layer of the epithelium-the basal layer. The intermediate layers lie upon the basal ...
caused by increased levels of
estrogen Estrogen or oestrogen is a category of sex hormone responsible for the development and regulation of the female reproductive system and secondary sex characteristics. There are three major endogenous estrogens that have estrogenic hormonal acti ...
. The cells may float in basophilic
mucous Mucus ( ) is a slippery aqueous secretion produced by, and covering, mucous membranes. It is typically produced from cells found in mucous glands, although it may also originate from mixed glands, which contain both serous and mucous cells. It is ...
. * Late-proestrus - there is a decrease in neutrophils because they are unable to cross the thickening vaginal wall. Cytology consists mostly of large intermediate and superficial cells. Red blood cells and
bacteria Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria were among ...
may be present.


Estrus

Estrus The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous p ...
is characterized by the presence of all superficial cells, many of which are anuclear (without a nucleus) or have small, pyknotic nuclei, and red blood cells may be present, along with bacteria. No neutrophils are seen unless there is
inflammation Inflammation (from la, wikt:en:inflammatio#Latin, inflammatio) is part of the complex biological response of body tissues to harmful stimuli, such as pathogens, damaged cells, or Irritation, irritants, and is a protective response involving im ...
. There is usually no mucous in the background.


Diestrus

In
diestrus The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous ...
, superficial cells are replaced by parabasal and intermediate cells within one to two days of onset. Neutrophils are increased. Red blood cells may be absent or present. Bacteria is commonly observed. A smear made during late estrus to early diestrus may appear similar to one made in early- or mid-proestrus.


Anestrus

The final stage of estrous is
anestrus The estrous cycle (, originally ) is the set of recurring physiological changes that are induced by reproductive hormones in most mammalian therian females. Estrous cycles start after sexual maturity in females and are interrupted by anestrous ...
, which is characterized by predominantly non-cornified squamous epithelial cells, such as intermediate or parabasal cells. The cells are smaller, have basophilic cytoplasm, and large round nuclei. There may be some neutrophils, but no red blood cells. The overall cellularity is low.


See also

*
Canine reproduction Canine reproduction is the process of sexual reproduction in domestic dogs, wolves, coyotes and other canine species. Canine sexual anatomy and development Male reproductive system Erectile tissue As with all mammals, a dog's penis is made up of ...
*
Reproductive cycle In biology, a biological life cycle (or just life cycle or lifecycle when the biological context is clear) is a series of changes in form that an organism undergoes, returning to the starting state. "The concept is closely related to those of the ...
*
Cytopathology Cytopathology (from Greek , ''kytos'', "a hollow"; , ''pathos'', "fate, harm"; and , '' -logia'') is a branch of pathology that studies and diagnoses diseases on the cellular level. The discipline was founded by George Nicolas Papanicolaou in 1 ...


References

{{reflist Diagnostic obstetrics and gynaecology Veterinary procedures