ANIMAL DEVELOPMENT

PRACTICAL ARTICLES

VAGINAL SMEAR ON THE MOUSE

Nanda Syafira 1*

1
ICP of Biology Education, Group 3, Biology Education, Makassar State University, Indonesia.

Abstrak
Deskripsi apusan vagina pada mencit (mice) adalah proses pengumpulan sampel sel dan bahan
biologis dari vagina mencit untuk analisis mikroskopis atau eksperimen ilmiah. Apusan vagina tikus
sering dilakukan untuk tujuan penelitian ilmiah, mengamati morfologi sel vagina atau penelitian
terkait reproduksi dan perkembangan. Prosedur pengambilan spesimen vagina pada tikus
biasanya meliputi langkah-langkah sebagai berikut: Persiapan, tikus yang menjalani tes pap smear
biasanya akan dibius terlebih dahulu agar tetap tenang selama proses berlangsung. Selain itu,
peralatan dan bahan steril disiapkan untuk meminimalkan risiko kontaminasi. Sampel, dokter atau
peneliti akan menggunakan alat lunak seperti kapas steril untuk mengambil sampel dinding vagina
tikus". Usap akan digerakkan dengan lembut di sepanjang dinding vagina untuk mengumpulkan
sel dan lendir yang mungkin mengandung informasi penting. Contoh pengobatan, setelah sampel
diambil, swab akan dimasukkan ke dalam tabung reaksi atau wadah steril untuk dibawa ke
laboratorium atau untuk dianalisis lebih lanjut.

Kata kunci: Tikus; Memegang; Tikus; Vagina; Tikus

Abstract
Description vaginal smear in mice (mice) is the process of collecting samples of cells and
biological material from the vagina of mice for microscopic analysis or scientific experiments.
Mouse vaginal smears are often performed for scientific research purposes, observing the
morphology of vaginal cells or reproduction and development related research. The procedure for
taking vaginal specimens in rats usually includes the following steps: Preparation, mice that
undergo a pap smear test will usually be anesthetized first to stay calm during the process. In
addition, sterile equipment and materials are prepared to minimize the risk of contamination.
Sample, tthe doctor or researcher will use a soft tool such as a sterile cotton swab to take a sample
of the mouse''s vaginal wall. The swab will be moved gently along the walls of the vagina to collect
cells and mucus that may contain important information. Examples of treatment, after the sample is
taken, the swab will be placed in a sterile test tube or container to be taken to the laboratory or for
further analysis.

Keywords: Rats;Holding; Mice; Vaginals; Mouse

INTRODUCTION
Description vaginal smear in mice (mice) is the process of collecting samples of cells
and biological material from the vagina of mice for microscopic analysis or scientific
experiments. Mouse vaginal smears are often performed for scientific research purposes,
observing the morphology of vaginal cells or reproduction and development related research.
The procedure for taking vaginal specimens in rats usually includes the following steps:
Preparation, mice that undergo a pap smear test will usually be anesthetized first to stay calm
during the process. In addition, sterile equipment and materials are prepared to minimize the
risk of contamination. Sample, tthe doctor or researcher will use a soft tool such as a sterile
cotton swab to take a sample of the mouse's vaginal wall. The swab will be moved gently
along the walls of the vagina to collect cells and mucus that may contain important
information. Examples of treatment, after the sample is taken, the swab will be placed in a
sterile test tube or container to be taken to the laboratory or for further analysis. Laboratory
analysis, in the laboratory, the sample will be analyzed under a microscope to observe the
structure of cells, organisms or other morphological changes that may be present in the
mouse vagina. Results and explanation, the results of the analysis of Mouse vaginal swabs
can provide insight into reproductive status, health, or ongoing research. These results can be
used to identify various changes or related conditions in the vagina of mice. It is important to
remember that smear testing in mice is often carried out as part of certain scientific studies or
laboratory studies. If you have any concerns about your pet's health, it is best to consult an
experienced veterinarian for appropriate advice and treatment.

METHOD
The tools and materials used in this practicum are the first tools, namely a dropper
pipette, light microscope, 250 ml beaker, tissue paper, sterile cotton, object glass, and cover
glass. The ingredients are female mice, 0.9% physiological NaCl, methylene blue, distilled
water, tap water, and 70% alcohol.
The way it works is to prepare a new object glass and cover glass, then use a new
dropper pipette. Wipe the tip of the cheek with cotton containing 70% alcohol. Put 0.5 ml of
physiological NaCl into the pipette. Insert the tip of the dropper into the mouse's vaginal
opening to a depth of approximately ½ cm, then spray physiological NaCl into the vaginal
opening. Suck out the liquid again and then spray it again into the vaginal opening. Do this
repeatedly until the Na fluid becomes cloudy. Place one drop of the physiological NaCl liquid
on the slide. Drop the physiological NaCl solution with 1 drop of methylene blue. Leave for
5-10 minutes. Discard the excess methyl blue dye, then carefully dip it into a beaker
containing tap water. Carefully suck up any excess fluid using tissue paper. Cover the object
glass with a cover glass. Do it carefully. Observe the vaginal smear preparation using a
microscope. Observe vaginal smear cytology using low magnification and gradually observe
with greater magnification. Draw the results of your observations in the space provided.
Determine the phase of the estrous cycle of the vaginal smear that you observed.
RESULT
Table 1. Results of Practical Activities
No Documentation Information
.
1. Documentation of tools and materials Recognize the tools and materials used in
vaginal smears and their functions (Tools
and materials in the photo are good).

2. Documentation of vaginal smear Place the mice in the cage correctly

preparation

3. Prepare all tools and materials used

correctly

4. Prepare all the tools used correctly

5. Prepare all the materials used correctly

6. Documentation of vaginal smear Holding mice properly.

preparation

7. Hold the dropper correctly.

8. Properly sample the vaginal epithelium

9. Make a vaginal smear correctly

10. Use the microscope correctly

12. Correctly observe vaginal smear cytology

13. Draw objects correctly

DISCUSSION
The white rat (Rattus norvegicus) is widely used as an experimental animal in various
studies. The use of laboratory animals in health research is often done to test the suitability or
safety of a drug or for research related to disease. Certified white mice are expected to make
it easier for researchers to obtain laboratory animals that meet the required criteria. The
criteria necessary for researchers to identify white rats as experimental animals include:
dietary control, Health Control, marriage registration, type (strain), age, weight, sex, genetic
genealogy. The White Rat has several characteristics that are favored as research test animals,
including its ability to reproduce quickly, its size is larger than mice, and ease of maintenance
in large numbers. White rats have characteristics such as small heads, albinos, tails longer
than the body, fast growth, the ability to produce a lot of milk, good temperament, and
resistance to arsenic thyroxine.(Frianto et al., n.d.)
 The external genitalia of female BALB/c mice are in a state of estrus, characterized
by the opening of the vagina and pink, moist, and swollen tissues (Figure 1a). The Vagina
expands so that wrinkles arise around the vagina. The estrus phase is characterized by an
enlarged vagina with slightly swollen, moist tissue, and a lighter red color intensity than the
proestrus phase (Figure 1B). The metestrus stage is characterized by a vaginal opening that is
not dilated, not swollen, not moist, and contains remnants of white blood cells.(Sulastri et al.,
2014)
tidak (Choi dan Rothman, 1990). Fenilalanin hasil metabolisme aspartam
menghasilkan asam amino esensial yang dibutuhkan tubuh karena merupakan prekursor
tirosin dan neurotransmitter. Fenilalanin dalam tubuh dimetabolisme melalui dua cara,
pertama fenilalanin diubah menjadi tirosin di hati oleh enzim fenilalanin hidrolase (Caballero
dan Wurtman, 1988). Tirosin merupakan asam amino esensial yang dibutuhkan sel saraf di
otak sebagai neurotransmitter. Untuk dapat memenuhi sawar darah otak, tirosin harus
diberikan dengan resep Pengangkut Asam Amino Netral (NAAT). Jalur kedua adalah
fenilalanin yang masuk ke dalam tubuh akan berhubungan langsung dengan NAAT dan
masuk ke otak tanpa mengalami tirosin. Dan fenilalanin untuk berikatan pada tempat
pengikatan NAAT. Ini akan menjadi tirosin berbicara NAAT. Jika fenilalanin mendominasi
di otak dibandingkan dengan asam amino lainnya, fenilalanin akan menjadi neurotoksik
(Fernstrom, 1994; Stegink dkk., 1987).(Pertiwi & Ihsani, n.d.)
The estrous phase in mice repeats itself and forms a continuous cycle called the
estrous cycle. The estrous cycle consists of four successive phases, proestrus, estrus,
metaestrus, and estrus. Estrus is one of the phases of the estrous cycle. The estrus period in
rats is 9 to 20 hours and the estrus cycle lasts 4 to 6 days. Experimental extract the duration of
the estrous cycle was determined by microscopic observation of the cells of the vaginal smear
seen during the proestrus phase characterized by multinucleated epithelial cells. This stage
marks the emergence of desires. In the period of estrus, the cervical smear is characterized by
the formation of keratinized cells (blood cells) that reflect the large number of mitotic
divisions that occur in the vaginal mucosa. Towards the end of the estrus phase, the lumen of
the vagina is formed by degenerated nucleated keratinocytes. During menstruation, the
ovaries contract and contain the corpus luteum, which contains luteinizing cells and small
nucleated follicles. The diestrus phase is dominated by leukocytes and nucleated epithelial
cells begin to form (Byers, 2012). This estrous cycle can not only be done with a vaginal
smear, but can also be observed in several ways, namely vaginal cytological examination,
electrical impedance measurement, biochemical water analysis, and visual observation of the
external genitalia. The estrous cycle of rats is also present in non-primate mammals. The
estrous cycle of rats is related to the menstrual cycle of primates and humans (Campbell,
2011). The stages of the estrous cycle can be classified by the type and proportion of cells
present in the sample obtained. This stage of estrus has special characteristics and during
estrus ovulation occurs. The estrus phase is characterized by strange, nervous and undeniable
behavior when the male approaches the female.(Putri et al., 2023)
Check the estrus cycle before and afte using the tomato extract is taken through
basting method. First steps to take is a sample of Mouse vagina (Mus musculus). Use a wet
cotton swab then add 0.9% NaCl solution. The Vagina of an adult female rat is then rubbed
slow. Cotton wool is applied with a thin layer and in the same direction as above on glass
objects using techniques smearing. The resulting stain is dried by drying in the air for about 5
to 10 minutes. Results dry smear received methylene dye
The blue color is then allowed to stand for about 30 minutes. The preparation is washed off
during use
aquades(Bahri et al., 2021)

CONCLUSION
The importance of rat breeding techniques is to maintain the physical condition and
physiology of rats The reproductive cycle in female mice consists of 4 phases, namely,
estrus,metestrus, diestrus and proestrus. The cells of the mouse phage smear consist of
leukocytes and epithelial cells, where the epithelial cells are differentiated into horn cells and
nucleated epithelial cells. Vaginal smear cells indicate the phases of the estrous cycle. The
proestrus phase is characterized by the presence of normal epithelial cells, having nuclei.
Estrus Phase found Gore epithelial cells, large size, no nucleus. The metestrus phase is
characterized by Gore epithelial cells and leukocytes are found. While the diestrus phase is
the discovery of nucleated epithelial cells and there are leukocytes.

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