Name: compound administration iii
Uploaded: 2023-04-30T06:46:40.000Z
Duration: 12 min 34 s
Channel: JoVE

K12 Standards

new-york-state-science-learning-standards

New York State Science Learning Standards

High School

Life Science

Structure and Function

HS-LS1-2.

Compound Administration III

1. Intradermal administration
<ol>
	<li>Most intradermal injections are aqueous-based compounds. The solutions must be physiologically buffered to have a neutral pH, in order to avoid tissue necrosis at the injection site.</li>
	<li>The needle size range is 25-30 gauge, the smallest possible.</li>
	<li>The dose range per injection site is 50-100 µL. Injecting excessive volumes can result in necrosis at the injection site or leakage of the compound out of the site due to pressure.</li>
	<li>For accurate placement of the needle into the intradermal space, it is necessary to anesthetize both mice and rats. Inhalation anesthesia allows for a rapid induction and recovery; however, injectable anesthesia has the advantage of providing ample time for preparation of the area and performing the injection.1</li>
	<li>Remove the hair at the injection site through the use of a depilatory cream or by shaving the area.</li>
	<li>Thoroughly remove any residual depilatory cream or hair debris.</li>
	<li>Apply a topical antiseptic, such as an iodine solution, chlorhexidine solution, or alcohol.</li>
	<li>Administration procedures
	<ol>
		<li>Stretch the skin taut between the thumb and index finger. This provides stability to the skin when positioning the needle.</li>
		<li>Place the needle bevel up on the skin.</li>
		<li>Gently insert the needle into the skin between the epidermis and the dermis. Advance the needle just beyond the bevel.</li>
		<li>Inject the substance slowly. Injection of the compound will create a bleb, or small blister, in the skin.</li>
		<li>Pause after the injection to allow the skin to stretch and adjust before withdrawing the needle.</li>
	</ol>
	</li>
	<li>Precautions
	<ol>
		<li>Pulling back on the plunger is not necessary.</li>
		<li>If the needle is inserted into the subcutaneous space, no bleb is formed. Injecting too deeply results in a subcutaneous injection.</li>
		<li>Avoid blotting or wiping the area, as that can cause the compound to leak from the injection site.</li>
		<li>When performing multiple injections, space them so that the blebs do not overlap.</li>
	</ol>
	</li>
</ol>
<img alt="Animal model injection, syringe administration on anesthetized mouse for experimental research study." src="/files/ftp_upload/10215/10215fig1.jpg" data-altupdatedat="1761830439000" data-alt="Figure 1"> 
Figure 1. Intradermal injection in mice.
2. Intranasal administration
<ol>
	<li>Equipment
	<ol>
		<li>Use a micropipetting unit pipette that can be calibrated to deliver an accurate volume.</li>
		<li>Disposable pipette tips should be used to avoid cross-contamination. TB syringes, blunt needles, and flexible tubing can also be used for dosing.</li>
		<li>Total injection volumes for rats should not exceed 40-100 µL, and should be given in 6-10 µL drops. For mice, the maximum total volume is 24 µL, given in 3-4 µL drops.</li>
	</ol>
	</li>
	<li>Administration in conscious animals
	<ol>
		<li>Manual restraint in conscious animals requires that the head be relatively immobile so that the pipette tip or blunt needle can be placed close enough to the nares to deliver the compound, but not so close as to poke or lacerate the nasal tissue.</li>
		<li>Restrain the animal and hold it in a vertical position.</li>
		<li>Place a small drop of a liquid compound at the nasal opening. The animal should inhale the droplet.</li>
		<li>Administer an additional volume, alternating nares until the entire volume has been given.</li>
		<li>When administering larger volumes, it is important to not constrict the chest during restraint. Chest compression impedes the ability of the animal to take sufficiently deep breaths to draw the liquid into the bronchi and lungs.</li>
	</ol>
	</li>
</ol>
<img alt="Laboratory mouse oral dosing; pipette method; experimental pharmacology; subject feeding setup." src="/files/ftp_upload/10215/10215fig2.jpg" data-altupdatedat="1761830439000" data-alt="Figure 2"> 
Figure 2. Intranasal administration in conscious mice.
<ol start="3">
	<li>Administration in unconscious animals
	<ol>
		<li>Using inhalation anesthetics allows for an immobilized animal during the approach and delivery of the compound. This eliminates the possibility of the animal biting the dosing equipment, the loss of the compound due to jerking of the head, and injury to the animal's nasal tissue, eyes, or facial skin. The animal is also less likely to snort and spray the compound from the nares upon administration.</li>
		<li>Place the animal in the supine position. The position of the head influences the placement of the solution in the nares. It has been demonstrated that the ideal position for CNS delivery is with the animal supine; this allows for better absorption.2</li>
		<li>Administer half of the compound directly into one side of the nasal opening, timing it with inhalation. Then, turn the animal over.</li>
		<li>As above, administer the other half of the volume into the other nasal opening.</li>
	</ol>
	</li>
</ol>
<img alt="Laboratory mouse anesthesia procedure, syringe, gloved hands, animal research, experiment setup." src="/files/ftp_upload/10215/10215fig3.jpg" data-altupdatedat="1761830439000" data-alt="Figure 3"> 
Figure 3. Intranasal administration in unconscious mice.
3. Intracranial administration in neonatal mice and rats
<ol>
	<li>Mice or rats must be anesthetized for intracranial injections.</li>
</ol>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td colspan="2">Mouse</td>
			<td colspan="2">Rat</td>
		</tr>
		<tr>
			<td>Age (days)</td>
			<td>Needle gauge (g)</td>
			<td>Age (days)</td>
			<td>Needle gauge (g)</td>
		</tr>
		<tr>
			<td>0-7 </td>
			<td>29-30</td>
			<td>0-5</td>
			<td>27-29</td>
		</tr>
		<tr>
			<td>7-14</td>
			<td>27</td>
			<td>5-10</td>
			<td>25-27</td>
		</tr>
		<tr>
			<td>14-28</td>
			<td>25</td>
			<td>10-14</td>
			<td>25</td>
		</tr>
		<tr>
			<td>Age (days)</td>
			<td>Needle length (mm)</td>
			<td>Age (days)</td>
			<td>Needle length (mm)</td>
		</tr>
		<tr>
			<td>0-7 </td>
			<td>2</td>
			<td>0-4</td>
			<td>2-3</td>
		</tr>
		<tr>
			<td>7-14</td>
			<td>3</td>
			<td>4-7</td>
			<td>3</td>
		</tr>
		<tr>
			<td>14-21</td>
			<td>4</td>
			<td>7-10</td>
			<td>4</td>
		</tr>
		<tr>
			<td>21-28</td>
			<td>5</td>
			<td>10-14</td>
			<td>5</td>
		</tr>
		<tr>
			<td>Age (days)</td>
			<td>Volume (µL)</td>
			<td>Age (days)</td>
			<td>Volume (µL)</td>
		</tr>
		<tr>
			<td>0-5</td>
			<td>&lt;20</td>
			<td>1-3</td>
			<td>&lt;20</td>
		</tr>
		<tr>
			<td>6-20</td>
			<td>&lt;60</td>
			<td>4-10</td>
			<td>&lt;60</td>
		</tr>
		<tr>
			<td>20-28</td>
			<td>&lt;100</td>
			<td>11-14</td>
			<td>&lt;100</td>
		</tr>
	</tbody>
</table>
Table 1. Needle gauge, needle length and maximum volume of intracranial administration as per the age of mice and rats.4
<ol start="2">
	<li>Equipment
	<ol>
		<li>Determine the correct needle gauge and maximum volume of administration as per Table 1.</li>
		<li>Prepare a needle guard prior to anesthetizing the animal. 
		NOTE: The injection depth of the needle is controlled through the use of a guard that is created with the needle cap.
		<ol>
			<li>To create the guard, a needle is measured against the needle cap, and a mark is placed on the cap to indicate where to cut. The cut should be made so that 2 to 5 mm of the needle are exposed when the cap is replaced on the needle.</li>
			<li>The length of the exposed needle must be long enough to penetrate the skin and skull, and reach the desired depth in the brain.</li>
			<li>The needle lengths needed for mice and rats are listed in Table 1.</li>
		</ol>
		</li>
		<li>A heating source is required to prevent hypothermia in the pups. There are several types: an electric heating pad set on low, a circulating water blanket, or a reusable chemical reaction heat pouch.</li>
	</ol>
	</li>
	<li>Restraint
	<ol>
		<li>Mouse and rat pups 10 days of age or younger do not require anesthesia for this procedure. Restrain them manually by holding them just behind the head and pulling the skin caudally to put slight pressure over the shoulders.</li>
		<li>Anesthetize pups over 10 days of age with isoflurane inhalation. Attach an induction chamber to a precision vaporizer or a bell jar with a cotton ball soaked with isoflurane. Once the pup is immobilized, the anesthesia is effective for about 40 seconds, which provides ample time for the injection.</li>
	</ol>
	</li>
	<li>Injection technique
	<ol>
		<li>Draw the substance into the syringe and place the cap/needle guide over the needle.</li>
		<li>The injection volume recommended as the maximum volume per neonatal mouse or rat is 100 µL, and that for weanling or older mice is up to 300 µL.</li>
		<li>To inject into the cerebral cortex of neonates, insert the needle 5 mm behind the eye, approximately 3 mm off the midline of the skull.</li>
		<li>The weanling mouse injection site is approximately half way between the eye and ear, and just off the midline.</li>
	</ol>
	</li>
</ol>
<img alt="Mouse injection experiment; syringe injecting substance; laboratory animal research; safety gloves." src="/files/ftp_upload/10215/10215fig4.jpg" data-altupdatedat="1761830439000" data-alt="Figure 4"> 
Figure 4. Intracranial administration in a mouse pup.

The administration of compounds into animals can have a significant effect on both the wellbeing of the animal and the outcome of the experimental data and scientific value. The proper method of delivery is essential to the success of the experiment. Many factors must be considered to determine the best route, including the scientific aim of the study, the pH of the substance, the required dosage volume, the viscosity of the substance, and the wellbeing of the animals. Technical expertise is also a requirement for all in...

compound administration iii

Source: Kay Stewart, RVT, RLATG, CMAR; Valerie A. Schroeder, RVT, RLATG. University of Notre Dame, IN
There are many commonly used routes for compound administration in laboratory mice and rats. However, certain protocols may require the use of less commonly used routes, including intradermal, intranasal, and intracranial injections. Specialized training is essential for these procedures to be performed successfully. Justification for these routes may need to be provided to gain Institutional Animal Care and Use Committee (IACUC) approval.

Source: Kay Stewart, RVT, RLATG, CMAR; Valerie A. Schroeder, RVT, RLATG. University of Notre Dame, IN
There are many commonly used routes for compound ...

At times, different experimental approaches necessitate the use of less commonly employed routes of compound administration in rodents. Intradermal, intranasal, and intracranial are three of such alternate routes that the biomedical researchers use in labs today.As their name suggests, intradermal is delivering compounds into the outer layers of the dermis. Intranasal is placing the solution in the animal's nares. And intracranial involves inserting the needle directly into the rodent's brain.Specialized training is essential to perform these procedures successfully. Here, we'll first illustrate the considerations for each of these methods and then we will demonstrate the techniques that will help you learn the procedures while ensuring safety of the animal and success of the experiment.Let's begin with the discussion of when these routes are usually employed and things one should bear in mind before beginning to perform these specialized administration techniques.The intradermal injections are used to deliver an article into the space between the epidermis and the dermis This route is usually reserved for the assessment of inflammation, cutaneous blood flow diagnostics, or allergenic reactions to an antigen. Similar to other routes, the intradermal solution should also be prepared using the sterile technique. And it must be physiologically buffered to have a neutral pH in order to avoid tissue necrosis at the injection site. A hub-less system with a 25-30 gauge needle is often used for this injection. This system helps in preserving the volume of administration, which is in the range of 50-100 microliters per injection site. Injecting excess can result in necrosis or undesirable compound leakage due to pressure.Intranasal route is often chosen for local delivery of vaccinations or decongestant spray as well as systemic and CNS delivery. The mucosa that lines the nasal cavity has a rich supply of blood vessels and nerves that allow for rapid systemic absorption and direct targeting to the CNS. This is a non-invasive method that requires minimal training and skill, and simple equipment - a calibrated micropipette and some disposable tips. The administration volumes for rats should not exceed 40-100 microliters given in 6-10 microliter-drops. And for mice, the maximum total volume is 24 microliters given in 3-4 microliter-drops.Although anesthesia is not required for this procedure, it has some advantages over intranasal administration in conscious animals 1) it facilitates proper placement of the compound at the nares, ensuring accurate dosing 2) eliminates the possibility of the animal biting the dosing equipment 3) ensures that there is no injury to the animal's nasal tissue, eyes, or facial skin due to jerking of the head, and 4) animal is less likely to snort and spray the compound from the nares upon administration.Intracranial injections in adult mice and rats employ the use of stereotaxic equipment, which is described in a video in the "Essentials of Neuroscience" collection. The equipment assures proper positioning and correct depth of injection. Here, we will focus on intracranial delivery in neonatal mice and rats in whom the skull is thin enough to inject directly through it, and may be too fragile to support the stereotaxic device. The primary purposes of this technique are to deliver CNS pharmacologic agents directly into to the CNS, and to avoid the effects encountered via any systemic route. The needle gauge, length and volume of administration are determined based on the species and the age of the pups. Note that as the animal's age increases, the gauge number decreases, the required needle length increases, and the maximum recommended administration volume also increases.With this background information in mind, let's delve into the procedures of these injection methods. First up is the intradermal administration technique. This procedure has to be performed in anesthetized animals. Review another video in this collection to understand the procedures for anesthesia induction and maintenance.Once the animal is anesthetized, shave the injection site using an electric razor or a depilatory cream. With a water-dampened gauze, thoroughly remove the lingering hair from the site. Then, with another gauze pad, apply a topical antiseptic solution to the shaved area. For administration, first stabilize the skin at the injection site by stretching it between your thumb and index finger.Now place the needle bevel up on the skin and gently insert it just beyond the bevel so that the opening is between the epidermis and the dermis layers. Then inject slowly and note that it creates a bleb in the skin. If the needle is inserted too deeply then no bleb will be formed. After injecting, pause to allow the skin to stretch and adjust, and then withdraw the needle slowly. Do not pull back on the plunger at anytime, as you would draw up the tissue and cause trauma at the injection site. Also, do not wipe or blot the injection site, as this may cause the injected substance to leak. When performing multiple injections, be sure to space them apart wide enough so that the blebs do not overlap with one another.Next, let's learn the intranasal administration procedure in conscious and anesthetized animals.For awake animals, restrain them by scruffing the skin at the nape of the neck and then hold the animal in a vertical position with its head immobilized. Be careful not to constrict the chest as that might impede the animal's ability to take sufficiently deep breaths to draw the liquid into the lungs. Using a micropipette, administer part of the solution by placing a small drop of liquid at the nasal opening. The animal will inhale the droplet. Repeat this process, alternating between the two nasal openings until the entire volume to be administered has been given. Recall - the total volume of administration should not exceed 24 ?l and 100 ?l in mice and rats, respectively.For anesthetized mice and rats, place the animal in a dorsal recumbency position. This position is ideal for CNS delivery as it allows for better absorption of the compound. Rotate the animal's head and administer half of the compound directly into one side of the nasal opening, timing it with inhalation. Then, rotate the animal's head to position for next administration. After 2 breaths or so, administer the remaining volume into the second nasal opening. After complete administration, return the animal back to its cage.Next, let's review the intracranial administration procedure for neonatal mice and rats. Prior to beginning the procedure, place the cage with the pups and the dam on an electric heating pad set to low. Make sure that a part of the cage is off of the heating pad. This is to prevent hypothermia and, at the same time, allow the dam to move away from the heat if she desires. Next, select a needle gauge appropriate for the age of the animal. Recall, the needle gauge; needle length, which is used to control the depth of the needle during the intracranial injection; and the volume of administration...all vary with animal's age and species.The length is adjusted using a guard. To prepare this guard, measure the correct needle against its cap and make a mark. Next, place a second mark on the cap to indicate where it will be cut. The distance between the two marks is the desired needle length. Then, cut the cap with a razor blade. Do not use scissors as they will crush the cap and will not produce a clean level cut. This is the "needle guard." Dispose the needle used for creating the guard, as it is no longer sterile, and instead insert a new needle into the guard and ensure that the correct length is exposed. Next, using a different needle attached to the suitable syringe, draw the injection substance. A different needle is used to do this, because placement into the stopper will significantly dull these fine gauge needles, which is not ideal for intracranial administration. Then, place the filled syringe on the needle with the guard. Now the system is ready for an injection.For pups over 10 days of age, administer inhalation anesthesia. Pups younger than 10 days old need not be anesthetized. To perform the injection, first locate the site, which is 5 mm behind the eye and approximately 3 mm off the midline of the skull. Next, insert the needle to the depth allowed for by the needle guard. Then, inject in a slow steady manner to avoid trauma to the brain. Remove the needle immediately and with great care to prevent injury to the brain tissue. Finally, place the animal back in with the dam to allow for proper recovery.Now let's review some experiments being conducted in labs today that utilize these uncommon routes of administration.An intradermal injection is often used to study skin inflammatory reaction. In this experiment, the researchers used this method to inject an allergen into one ear and a neutral substance into the opposite ear of a pre-sensitized mouse. Next, they delivered a blue dye into the animal's circulatory system to examine the changes in vascular permeability due to allergen injection.As mentioned earlier, one of the applications of intranasal administration is to administer vaccines. Here, scientists used this route to deliver a genetically modified, live attenuated influenza vaccine into the wild type and transgenic mice and studied mucosal immunity via production of a specific type of T-cells.Lastly, these biomedical researches used intracranial administration to implant cancer cells in immunocompromised mice, in order to create a human brain tumor model. The efficacy of the injection was then analyzed using an in vivo imaging system.You have jus watched JoVE's video on some of the special methods of compound administration in laboratory mice and rats. You should now understand when these procedures are helpful, the considerations you should bear in mind before and while performing these techniques, and the essential procedural steps to ensure that the administration has minimal impact on the animal's health and on the experimental data to be collected. As always, thanks for watching!

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lab animal research

This collection is a comprehensive video guide for appropriate lab animal care and use. Since a majority of biomedical research is focused on studies involving rodents, it is critical that every scientist learns the essential procedures demonstrated in these videos.

Learn why intradermal, intranasal, and intracranial injections may be used in laboratory mice and rats, and why training and IACUC approval matter.

Specialized Injection Routes in Mice and Rats | JoVE K12

HS-LS1-3.

Kay Stewart

Valerie A. Schroeder

University of Notre Dame

Overview

Considerations for the Specialized Injections

Intradermal Administration

Intranasal Administration

Intracranial Administration in Neonatal Rodents

Applications

Summary

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Community DNA Extraction from Bacterial Colonies

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - The University of Arizona
Demonstrating Author: Luisa Ikner
Traditional methods of analysis ...

Cytogenetics

Cytogenetics is the field of study devoted to chromosomes, and involves the direct observation of a cell&#8217;s chromosomal number and structure, ...

embryonic-stem-cell-culture-and-differentiation

Embryonic Stem Cell Culture and Differentiation

Culturing embryonic stem (ES) cells requires conditions that maintain these cells in an undifferentiated state to preserve their capacity for self-renewal ...

DNA Gel Electrophoresis

DNA gel electrophoresis is a technique used for the detection and separation of DNA molecules. An electric field is applied to a gel matrix comprised of ...

Enzyme Assays and Kinetics

Enzyme kinetics describes the catalytic effects of enzymes, which are biomolecules that facilitate chemical reactions necessary for living organisms. ...

DNA Ligation Reactions

In molecular biology, ligation refers to the joining of two DNA fragments through the formation of a phosphodiester bond.  An enzyme known as a ligase ...

explant-culture-for-developmental-studies

Explant Culture for Developmental Studies

Explant culture is a technique in which living cells or tissues are removed from an embryo for continued development outside of the organism. This ex vivo ...

DNA Methylation Analysis

Methylation at CpG dinucleotides is a chemical modification of DNA hypothesized to play important roles in regulating gene expression. In particular, the ...

Expression Profiling with Microarrays

Microarrays are important tools for profiling gene expression, and are based on complementary binding between probes that are attached to glass chips and ...

Density Gradient Ultracentrifugation

Density gradient ultracentrifugation is a common technique used to isolate and purify biomolecules and cell structures. This technique exploits the fact ...

F&#246;rster Resonance Energy Transfer (FRET)

F&#246;rster resonance energy transfer (FRET) is a phenomenon used to investigate close-range biochemical interactions. In FRET, a donor photoluminescent ...

detecting-environmental-microorganisms-with-polymerase-chain-reaction

Detecting Environmental Microorganisms with the Polymerase Chain Reaction and Gel Electrophoresis

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University
Demonstrating Author: Bradley Schmitz
Polymerase chain reaction (PCR) is ...

Gel Purification

Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, ...

Detecting Reactive Oxygen Species

Reactive oxygen species are chemically active, oxygen-derived molecules capable of oxidizing other molecules. Because of their reactive nature, there are ...

Gene Silencing with Morpholinos

Morpholino-mediated gene silencing is a common technique used to study roles of specific genes during development. Morpholinos inhibit gene expression by ...

development-and-reproduction-of-the-laboratory-mouse

Development and Reproduction of the Laboratory Mouse

Successful breeding of the laboratory mouse (Mus musculus) is critical to the establishment and maintenance of a productive animal colony. Additionally, ...

Genetic Crosses

To dissect genetic processes or create organisms with novel suites of traits, scientists can perform genetic crosses, or the purposeful mating of two ...

drosophila-melanogaster-embryo-and-larva-harvesting-and-preparation

Drosophila melanogaster Embryo and Larva Harvesting and Preparation

Drosophila melanogaster embryos and larvae are easy to manipulate and develop rapidly by mechanisms that are analogous to other organisms, including ...

Genetic Engineering of Model Organisms

Transgenesis, or the use of genetic engineering to alter gene expression, is widely used in the field of developmental biology. Scientists use a number of ...

electrophoretic-mobility-shift-assay-emsa

Electrophoretic Mobility Shift Assay (EMSA)

The electrophoretic mobility shift assay (EMSA) is a biochemical procedure used to elucidate binding between proteins and nucleic acids. In this assay a ...

Construct an explanation based on evidence for how the structure of DNA determines the structure of proteins which carry out the essential functions of life through systems of specialized cells.

HS-LS1-1.

Compound Administration III

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