IACUC Protocol Submission Checklist
For deadline submission dates please click on the ‘IACUC’ tab on the Animal
Facility Website: http://squeek.eri.harvard.edu
Before submission to IACUC office for Pre-Review:
Pat D’Amore
 Pre-Approval by the DOR is required for all animal research that has not been
peer reviewed and must be obtained prior to submitting the protocol to the
IACUC office. The DOR Pre-Approval Form can be found on the IACUC
website. Please complete and submit this form to Pat D’Amore, allowing up to
7 days for review and approval.
 The one page Lay summary submitted for DOR pre-approval should be cut and
pasted into Section A2 of the actual protocol submitted to the IACUC office.
Kathleen Gallagher
 If needed, are SASPs and/or COMS attached to the protocol
 The use of biohazards, chemical hazards, radioactive material or physical
hazards (i.e. lasers), requires Kathleen Gallagher to review the protocol.
(SECTIONS D1gi, D1gii or APPENDIX A)
Reviewed by Kathleen Gallagher
Marie Ortega
 If the animals require special care/special housing, use of the Satellite facility,
BL2 use or any animal facility staff assistance, Marie Ortega must review the
protocol. (SECTIONS C2, D1giii or D4).
Reviewed by Marie Ortega
Karen Krueger
 Any questions regarding surgical procedures or the use of anesthetics/analgesics
should be answered by the veterinarian.
Review by the Attending Veterinarian, Karen Krueger
Rodent Pathogen Testing
 Are rodent pathogen testing results attached (the use of rodent derived materials
(i.e. serum, antibodies, cells)
Pre-Review of Protocol:
1
 Submitted to the IACUC Coordinator
 Before final submission of protocol:
o Obtain all signatures (PI, Kathleen Gallagher and Marie Ortega)
o Submit 1 hard copy to the IACUC Coordinator
Contacts
Candace Beiler, IACUC Coordinator ext. 408 acuc@schepens.harvard.edu
Meredith Gregory-Ksander, IACUC Chairman ext. 455
meredith_gregory@meei.harvard.edu
Marie Ortega, Associate Director of Animal Facility ext. 441
marie_ortega@meei.harvard.edu
Karen Krueger, Attending Veterinarian Karen.krueger@schepens.harvard.edu
Kathleen Gallagher, Health and Safety Officer ext. 244
kathleen_gallagher@meei.harvard.edu
Revised 6/18/2013
PI:
Protocol Number:
SCHEPENS EYE RESEARCH INSTITUTE
Institutional Animal Care And Use Committee
Animal Studies Protocol
The Schepens IACUC requires the following information to comply with the “Public Health Service Policy on
Humane Care and Use of Laboratory Animals”, the ILAR Guide, and the “Animal Welfare Act.” Address each item
independently, without reliance on information covered in other sections. Address only the questions asked, and do
not submit major sections of grant proposals or excessive details of assays not related to the use of live animals e.g.,
biochemical and/or molecular biology assays, and in vitro tests.
____________________________________________________________________________________________
This box for IACUC Office Use Only:
Protocol #:
Approval Stamp:
Termination Date:
SECTION A
A1. ADMINISTRATIVE INFORMATION
PI Last Name:
First Name:
Degree:
Title:
Primary Working Investigators:
Technician/Student:
Emergency contact name and No. (e.g. cell phone, pager or home telephone):
*Regardless of the actual project start and termination dates, the IACUC protocol will be considered approved for 3
years, starting with the date listed in the approval letter, after 3 years, a submission of a new protocol is required.
Title of Project (include species and procedure in title (no abbreviations)):
New Project
Renewal
2
A2. STUDY OBJECTIVES:
Since not all reviewers are familiar with your area of research,
NON-SCIENTIFIC TERMS MUST BE USED
A. Nontechnical description of the project and its potential value (limit to 1 page): In the space
provided below briefly describe the overall purpose, goals and significance (importance to the
advancement of scientific knowledge, general contribution to the well-being of mankind and/or
potential benefits for amelioration of disease or suffering) of your project. Bearing in mind that the
IACUC membership also includes administrative personnel, and community volunteers, use language
understandable to that of a 12th grade high school student level. Descriptions with excessive jargon,
scientific terminology, laboratory procedures, and/or grant specific aims will be not be accepted.
• Overall purpose:
• Goals:
• Significance:
• How this protocol and the use of animals will accomplish these goals:
B. Must provide glossary of technical terms used in protocol. Definitions must be in lay terms.
C. Renewal of previous study
If this Animal Studies protocol is a renewal of an expiring protocol, please provide a brief description
of your findings, and list any publications or presentations that may have resulted from your work.
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PI:
Protocol Number:
3
SECTION B
B1. RATIONALE FOR ANIMAL USE
a. Assurance of Unnecessary Duplication.
This study is peer-reviewed (e.g. NIH, competitive internal peer review).
Please be sure that the grant noted below is active and/or has been reviewed and received a
“fundable score”.
Granting Agency:
Grant #:
Grant Title:
This study will not be peer reviewed. Please submit the signed DOR Pre-Approval for NonPeer Reviewed Animal Research form with the protocol (available on the IACUC
website).
Give the following assurance that the activities do not unnecessarily duplicate any
previous efforts:
Database Used (PubMed, Reporter)
Years
Covered
Key Words Used
Date of
Search
Search
Results
b. Key references supporting this study (full references, 5 maximum).
c. What are the possible alternatives to animal use and why were they rejected?
d. Why is this species and these strains (if applicable) used in the experiments? Please
list all strains (i.e., transgenics and knock-outs) as well as include a brief rationale.
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PI:
Protocol Number:
4
SECTION C
C1. DESCRIPTION OF EXPERIMENTAL DESIGN AND JUSTIFICATION FOR ANIMAL
NUMBERS
When designing your experiments, please limit animal involvement by using the minimum number
required to obtain reliable results and consider the use of non-animal methods, such as mathematical
models, computer simulation, or in vitro biological systems when possible. (for example see
APPENDIX G)
Each of the following MUST be addressed in your response:
- Rationale for each experiment (Hypothesis, question being asked, how the proposed
experiment will answer the question)
- Define the groups and number of animals per group needed for each experiment (including
both experimental and control animals).
- Justify how the number of animals for each experiment was determined (statistics, power
analysis, previous publications, amount of tissue needed etc.)
NOTE: All animals involved in this proposal must be included and justified in this section.
This includes breeding pairs, pregnant mothers, pups used before weaning, and offspring that cannot
be used.
A table/flow chart describing experimental groups and controls MUST be provided.
(for example see APPENDIX I)
SUMMARY TABLE OF ALL ANIMALS:
Species/Strain
Sex, Age, Weight
Total # Animals
Total Number of Animals to be Used
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PI:
Protocol Number:
5
C2. GENETICALLY MODIFIED ANIMALS (BY MANIPULATION OR SPONTANEOUS
MUTATION):
Describe any phenotypic consequences of the genetic manipulations to the animals. List each strain
and describe any special care or monitoring that the animals will require (e.g., autoclaving of cages,
special diet, specialized caging or environment, etc) or monitoring that the animals will require. If
any special care or monitoring requires animal facility staff assistance, preapproval by the
animal facility manager, or designate, must be obtained before submitting the protocol (see
Section I3).
Strain
Revised 6/18/2013
Phenotypic Consequences
Special Care
PI:
Protocol Number:
6
SECTION D.
D1a. PROCEDURE CHECKLIST (Please provide detailed description in Section D1b.)
Please check each procedure proposed in this protocol. Please note this list is for live
animals only.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
Activity wheel
Anesthesia by gas
Anesthesia by injectibles
Aqueous humor withdraw
Breeding
Cardiac blood collection
Corneal wounding
Craniotomy
Diabetes Induction
Dry eye chamber
Enucleation (survival)
Electroporation (in vivo)
Electroretinogram (ERG)
Euthanasia by CO2
Euthanasia by drugs
Experimental autoimmune disease
(EAU)
Fluorescein angiogram
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
Fluorescein Corneal Staining
Food restriction
Fundus exam
Gavage
Genotyping
HRT
ID
Immunization with adjuvant
Immunization without adjuvant
Implants
Inflammatory immunoassay
Injection of biohazards (skin, iv,
im, ip, not ocular)
Injection of cells (skin, iv, im, ip,
not ocular)
Injection of chemicals (skin, iv,
im, ip, not ocular)
IOP
Irradiation exposure
Lacrimal gland excision
Monoclonal antibody production
(ascites)
53
54
55
56
57
58
59
60
61
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52
Neurotoxin injections
OCT
Ocular injection of biohazards
Ocular injection of cells
Ocular injection of chemicals
Optic nerve damage
Orbital injection
Ovariectomies – SX
Oxygen chamber
Perfusion
Retinal laser burn
Retro-orbital blood collection
Splenectomy
Tarsorrhaphy
Thioglycolate IP injections
Topical (Cornea) Application of
biohazards
Topical (Cornea) Application of
chemicals
Training
Tran limbal laser photocoagulation
Transcleral incision
Transplantation of cells
Transplantation of organs
Transplantation of tissues
Traumatic brain injury
Tumor growth
Other:
PI:
Protocol Number:
7
D1b. DESCRIPTION OF ANIMAL PROCEDURES.
- Provide the sequence and timing of all live animal procedures to be performed.
A timeline, diagram, or flowchart must be used. If proposing multiple experimental sets, please
provide a separate timeline for each experimental set as described in Section C1. (For example see
APPENDIX J) This may be included within the text box below or as a separate document.
- Provide a brief description of each procedure involving live animals (Surgical and Non-Surgical).
(For example see APPENDIX H)
NOTE: IACUC reviewer should be able to understand exactly what will be done to all of the animals
from the entry into the study to the endpoint of the study.
Please go to the link below to review SASP/LSP#000 Safe Laboratory Practices. The PI must
initial the box provided which confirms that you acknowledge and will follow SASP/LSP#000
when performing the procedures stated within this protocol.
http://squeek.eri.harvard.edu/health_safety.html
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PI:
Protocol Number:
8
c. POST ANESTHESIA RECOVERY & MONITORING
Please check all that apply for immediate post anesthesia recovery and monitoring.
No post operative care is necessary – surgery is Non-Survival
Ophthalmic ointment will be applied to both eyes
Analgesics and other therapeutic agents will be provided as described in the protocol
Animals/cage will be placed on a water heat blanket in the procedure area until animals are awake and
sternal (able to maintain an upright position)
Animals will be monitored every 15 minutes to ensure they are breathing and they appear comfortable,
both in position and demeanor. Findings will be documented on the Cage Clinical Record located on the
back of their cage card
Animals will be monitored until they are sternal (able to maintain an upright position)
Animals/cage will be returned to the housing room after food and water are checked – animals are
awake and sternal
Cage Clinical Record will be filled out legibly and Procedure cage Flag will be placed on the cage
Other:
Any procedure on a live animal in which an incision is made is to be considered a surgical
procedure.
Will a surgical procedure be performed on animals (survival & non-survival)?
yes, you MUST complete Appendix B
Revised 6/18/2013
Yes
PI:
Protocol Number:
No. If
9
d. Therapeutic agents: Anesthetics, analgesics, antibiotics, ophthalmics
Procedure
Species, Agent, Dose(mg/kg body weight)
Route of
Admin.
Needle
Gauge
Frequency and Duration
Pharmaceutical
Grade*
If ‘No’, please fill
out Appendix C
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
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PI:
Protocol Number:
10
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
Mice: N/A
Rats: N/A
Rabbits: N/A
Other:
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
*All anesthetics, analgesics and ophthalmic antibiotic or lubricating ointments and eye drops MUST be pharmaceutical grade. These
agents are of paramount importance during a study and provide animals with minimal untoward side effects or toxicity. A list of
these compounds is located in the Appendix section of this protocol and is available from the Animal Facility. Use of an agent not
listed, requires approval by the IACUC.
The Policy for Non-Pharmaceutical Grade Compound Use can be found at: http://squeek.eri.harvard.edu/iacuc.html
Revised 6/18/2013
PI:
Protocol Number:
11
e.
Non-toxic experimental agents (For hazardous agents see Section D1g)
Will experimental agents be administered to animals that are NOT toxic, carcinogens,
biohazards, anesthetizing, or infectious as part of the protocol (by gavage, injection, topical
application, or some other method)?
Yes
No. If yes, please fill out table below.
Agent(s)
Procedure
Volume and
Frequency
Anesthetic, Pharmaceutical
if
Grade*
Concentration,
applicable
If ‘No’, please
if applicable
fill out
Appendix C
N/A
N/A
N/A
N/A
N/A
The Policy for Non-Pharmaceutical Grade Compound Use can be found at:
http://squeek.eri.harvard.edu/iacuc.html
If needed, describe in more detail what agent(s) is to be administered, how it is to be administered, etc:
f.
Non-hazardous, biological materials/animal products (For hazardous agents see Section D1g)
Will any biological material/animal products be used in animals (e.g., antibodies, cell lines, etc.)?
Yes
No. If yes, please list and describe materials (include species source & vendor).
Please note whether the agents are pharmaceutical grade or non-pharmaceutical grade. If the
agents are non-pharmaceutical grade, please complete Appendix C.
The Policy for Non-Pharmaceutical Grade Compound Use can be found at:
http://squeek.eri.harvard.edu/iacuc.html
Is material sterile or attenuated?
Yes
No
Other. Describe:
If tumor cell lines are to be used in rodents, please indicate the following below: (1) where and how the
cell line is implanted (2) describe in more detail how large the tumor will be allowed to grow.
Revised 6/18/2013
PI:
Protocol Number:
12
If cell lines or materials described above are derived from rodents, or grown in rodent serum, has the
material been tested for rodent pathogens?
See Animal Facility Website for more information: http://squeek.eri.harvard.edu/about.html
Yes (Attach copy of results)
No (contact veterinary staff for testing suggestions
g. Hazardous materials and lasers (radioisotopes, biohazardous biologicals, chemical agents,
lasers).
i)
Are test substances administered to animals that may be mutagens, radioactive, toxic,
carcinogens, bio-hazardous or infectious as part of the protocol?
Yes
No.
If yes, you MUST complete Appendix A and obtain a signature from the SERI
Safety Officer. If you are not sure, please see the SERI Safety Officer.
ii)
Will any procedures be performed on animals involving the use of physical hazards (i.e.
lasers, UV light).
Yes
No If yes, you MUST complete Appendix A and obtain a signature from
the SERI Safety Officer. If you are not sure, please see the SERI Safety Officer.
iii)
Will the animals need to be housed in the Animal Biosafety Level 2 Facility (ABL2)?
Yes
No If yes, pre-approval from the animal facility office MUST be
obtained before submitting the protocol. (See Section I2)
h. Tissue and Fluid Collection
Are specimens (e.g., blood, tissue for genotyping or other body fluid) collected non-surgically
from live animals?
Yes
No. If yes, please fill out table below.
If tail clipping animals, please state the age that the animals will be when the procedure is
performed.
Specimen
Procedure
Volume, if
applicable
Frequency
Anesthetic, if
applicable
N/A
N/A
If needed, describe in more detail what specimen is to be collected, how it is to be collected, etc:
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PI:
Protocol Number:
13
i.
Potentially painful/distressful procedures
Will animals be subject to experimental procedures that have the potential to cause
pain/distress other than those described above (e.g., exposure to noxious stimuli, food/water
deprivation, immunization, physical restraint >5 minutes, animal models of infectious disease, animal
models with potential physical consequences such as diabetes, etc.?)
Yes
No
If yes, please describe in more detail below. Include how animals will be monitored, and for what
clinical signs.
If unrelieved pain/distress (Category E), scientific justification must be provided in Section E2
j.
Identification methods
Describe individual animal identification methods (e.g., ear tags, tattoos, cage card, etc.) Please refer
to SERI’s IACUC Rodent ID Policy.
k. Eye Studies
Will both eyes be studied in this protocol?
Yes
No
If Yes, please describe each procedure that will be performed on both eyes.
Do any of these procedures have the potential to disrupt vision bilaterally?
Yes
No
If Yes, please scientifically justify the need to use both eyes.
Protocols involving bilateral survival ocular procedures require special consideration and
justification, with particular attention to any visual consequences. Such procedures include bilateral
ocular surgeries, whether performed simultaneously or sequentially, and any experimental procedure
that results in, or has the potential to result in, a level of visual disability sufficient to disrupt an
animal’s normal daily activity. Such procedures include: bilateral induction of laser-induced
choroidal neovascularization, bilateral intravitreal or subretinal injections, and systemic
administration of compounds resulting in bilateral disease (endotoxin-induced uveitis, experimental
autoimmune uveitis, sodium iodate-induced RPE degeneration, etc.)
l.
Special Housing (dry eye chamber, high oxygen chamber, etc.) Describe in detail the special
housing unit and how temperature, humidity and lighting will be maintained. If staff assistance is
required please refer to Section D4.
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PI:
Protocol Number:
14
D2. EXPERIMENTAL ENDPOINT CRITERIA
List the experimental endpoints for all experimental groups. All animal use must have a defined end
point and death as an endpoint must always be scientifically justified.
D3. PREMATURE EUTHANASIA CRITERIA
List the criteria to be used to determine when premature euthanasia is to be performed. Tumor size,
percentage body weight gain or loss, inability to eat or drink, behavioral abnormalities, clinical
symptomatology, or signs of toxicity must be specified when the administration of tumor cells,
biologics, infectious agents, radiation or toxic chemicals are expected to cause clinical symptoms or
are potentially lethal.
D4. SPECIAL CARE.
Is animal facility staff assistance required for special housing, diet, drug treatment, or health checks?
Yes
No. If yes, please describe below.
If any procedure requires animal facility staff assistance or monitoring, pre-approval from the
animal facility office MUST be obtained before submitting the protocol. (See Section I2.)
Will live animals be removed from the basement facility of 20 Staniford Street building?
Yes
No
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PI:
Protocol Number:
15
If yes, will live and/or dead animals be returned to the basement facility of 20 Staniford
Street building?
Yes
No
Will animals be housed in SERI 3rd floor Satellite Facility?
Yes
No
If yes, please refer to the policy on Satellite Facility use on the Animal Facility website:
http://squeek.eri.harvard.edu/policies.html
Describe and justify removing live animals from the facility and/or use of Satellite Facility below:
SECTION E
E1. PAIN OR DISTRESS CATEGORY
Classification B
Animals being bred, conditioned, or held for use in teaching, testing, experiments, research, or
surgery, but not yet used for such purposes.
Classification C
Minimal, transient, or no pain or distress (ear snipping, tail clipping prior to 21 days old,
subcutaneous/intradermal/intraperitoneal injections, facial vein/tail vein blood collection, slit-lamp
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PI:
Protocol Number:
16
exams)
Classification D
Pain or distress relieved by appropriate measures (surgery with use of analgesics, tail clipping after
21 days old)
Classification E
Animals upon which teaching, experiments, research, surgery, or tests will be conducted involving
accompanying pain or distress to the animals and for which the use of appropriate anesthetic,
analgesic, or tranquilizing drugs will adversely affect the procedures, results, or interpretation of the
teaching, research, experiments, surgery, or tests.
Examples:

Procedures producing pain or distress unrelieved by analgesics such as toxicity studies,
microbial virulence testing, radiation sickness, and research on stress, shock or pain.

Surgical and postsurgical sequelae from invasion of body cavities, orthopedic
procedures, dentistry or other hard or soft tissue damage that produces unrelieved pain
or distress

Negative conditioning via electric shocks that would cause pain in humans.

Performing an experimental procedure that will induce blindness in both eyes
The IACUC is responsible for applying U.S. Government Principle IV: "Proper use of animals, including
the avoidance or minimization of discomfort, distress, and pain when consistent with sound scientific
practices, is imperative. Unless the contrary is established, investigators should consider that procedures
that cause pain or distress in human beings might cause pain or distress in other animals.” If any animals
are listed under Category E provide scientific justification for withholding anesthesia, analgesia or
tranquilizing agents on the next page.
Species
(Common name)
USDA Classification
B, C, D, E.
Total number of animals (should equal total from Section C)
3-Year total number
of animals
Total:
E2. SCIENTIFIC JUSTIFICATION FOR CATEGORY E
Describe procedures producing pain or distress in these animals and the scientific justification for
not using appropriate anesthetic, analgesic or tranquilizing drugs.
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PI:
Protocol Number:
17
SECTION F
F1. CONSIDERATION OF ALTERNATIVES TO PAINFUL AND/OR STRESSFUL
PROCEDURES.
If any procedures fall into Category D or E, causing more than momentary or slight pain or distress to the
animals, you must certify that no valid alternative was identified to any described procedures which may
cause more than momentary pain or distress, whether relieved or not. Describe your consideration of
alternative procedures, and your determination that alternatives are not available or not appropriate for
your study. Alternatives include methods that (1) refine existing tests by minimizing animal distress, (2)
reduce the number of animals necessary for an experiment, or (3) replace whole- animal use with in vitro
or other tests. When ascites production is used to produce antibodies, scientific justification needs to
be given as to why in vitro systems cannot be used.
This is NOT a search for justification of the research project, it is a search for alternatives to
painful procedures. See examples in Appendix K
Responses must include:
1.
Procedure:
2.
Keywords:
3.
Database searched:
4.
Date of the search:
5.
Period of time covered by the search: month/year – month/year
6.
The results of the search and describe your consideration of alternative procedures and
your determination that alternatives are not available.
For help with your literature search or additional databases contact the Animal Welfare
Information Center at www.nal.usda.gov/awic/databases/database.htm or (301) 504-6212 or
awic@nal.usda.gov.
Literature Search #1
Literature Search #2
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PI:
Protocol Number:
18
Literature Search #3
Literature Search #4
Literature Search #5
Revised 6/18/2013
PI:
Protocol Number:
19
SECTION G
G1. WHAT AGENTS AND METHODS OF EUTHANASIA WILL BE USED?
EUTHANASIA (Euthanasia must be conducted in accordance with the AVMA Guidelines on Euthanasia
Please refer to “Rodent Euthanasia Policy” http://squeek.eri.harvard.edu/iacuc.html
Indicate the method of euthanasia to be employed
Overdose of pentobarbital 100 mg/kg body weight i.p. for rodents (See Note A)
Overdose of pentobarbital 120 mg/kg body weight i.v. for rabbits and larger animals
(See Note A)
CO2 narcosis (See Note A)
Perfusion of anesthetized animal anesthetic to be used:
dose:
route of administration:
Exsanguination of anesthetized animal
anesthetic to be used:
dose:
route of administration:
Cervical dislocation of anesthetized mouse
anesthetic to be used:
dose:
route of administration:
Decapitation of an anesthetized rodent
anesthetic to be used:
dose:
route of administration:
Cervical dislocation w/out prior anesthesia
(CATEGORY E: provide scientific justification for withholding anesthesia Section E2)
Decapitation without prior anesthesia
(CATEGORY E: provide scientific justification for withholding anesthesia Section E2)
Decapitation of neonatal mice or rats
Other – provide a detailed description of the method used to achieve euthanasia:
Note A. Indicate what methods will be used to ensure that the animal is dead prior to collecting
tissues or carcass disposal. Check all (at least 2) that are applicable.
Palpation of heart to ensure cessation of breathing
Monitor respirations to ensure cessation of breathing
Check reflexes (palpebral, toe pinch) to ensure no response
Open chest cavity, following euthanasia
Cervical dislocation (mouse only)
Other:
SECTION H
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PI:
Protocol Number:
20
Individually list all investigators, faculty (co-investigators), fellows/post-docs, technicians and
students to be covered by this protocol. Please answer a - g for each person listed. Copy sections as
needed.
1. Principal Investigator
a) Name:
b) Is this person:
New to the lab
Current lab member
c) Responsibilities on this protocol (be specific):
d) Please describe any previous experience working with animals:
e) Please describe specific experience performing the animal procedures described in this
protocol:
f) When were they trained at Schepens. If not yet trained, when will training begin. (online
and hands-on):
g) If necessary, who will train them at Schepens on the procedures described in this
protocol:
Add additional Principal Investigators below. Please be sure to label (a-g).
2. Faculty (Co-Investigator)
a) Name:
b) Is this person:
New to the lab
Current lab member
c) Responsibilities on this protocol (be specific):
d) Please describe any previous experience working with animals:
e) Please describe specific experience performing the animal procedures described in this
protocol:
f) When were they trained at Schepens. If not yet trained, when will training begin. (online
and hands-on):
g) If necessary, who will train them at Schepens on the procedures described in this
protocol:
Add additional Faculty (Co-Investigator) below. Please be sure to label (a-g).
Revised 6/18/2013
PI:
Protocol Number:
21
3. Fellows/Post-docs
a) Name:
b) Is this person:
New to the lab
Current lab member
c) Responsibilities on this protocol (be specific):
d) Please describe any previous experience working with animals:
e) Please describe specific experience performing the animal procedures described in this
protocol:
f) When were they trained at Schepens. If not yet trained, when will training begin. (online
and hands-on):
g) If necessary, who will train them at Schepens on the procedures described in this
protocol:
Add additional Fellows/Post-docs below. Please be sure to label (a-g).
4. Technical Staff
a) Name:
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PI:
Protocol Number:
22
b) Is this person:
New to the lab
Current lab member
c) Responsibilities on this protocol (be specific):
d) Please describe any previous experience working with animals:
e) Please describe specific experience performing the animal procedures described in this
protocol:
f) When were they trained at Schepens. If not yet trained, when will training begin. (online
and hands-on):
g) If necessary, who will train them at Schepens on the procedures described in this
protocol:
Add additional Technical Staff below. Please be sure to label (a-g).
5. Students
a) Name:
b) Is this person:
New to the lab
Current lab member
c) Responsibilities on this protocol (be specific):
d) Please describe any previous experience working with animals:
e) Please describe specific experience performing the animal procedures described in this
protocol:
f) When were they trained at Schepens. If not yet trained, when will training begin. (online
and hands-on):
g) If necessary, who will train them at Schepens on the procedures described in this
protocol:
Add additional Students below. Please be sure to label (a-g).
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23
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SIGNATURE PAGE
I1. ASSURANCE OF THE PRINCIPAL INVESTIGATOR.
The principal investigator assures the Schepens Institutional Animal Care and Use Committee they
understand that:
1. Investigators, technicians and students on this project will use animals in full compliance with
the PHS Policies on humane care and use of laboratory animals, the Animal Welfare Act, the
“Guide for the Care and Use of Laboratory Animals”, and Schepens Eye Research Institute
policies governing the use of live vertebrate animals for research purposes, and that
noncompliance could result in a loss of NIH grant funding and access to the animal facility.
2. The IACUC approval is valid for a maximum of 36 months following the date of original
approval with annual reviews, and that the protocol is under constant evaluation for
compliance.
3. An Amendment Form will be submitted to the IACUC (or person designated by the IACUC)
for committee approval of minor changes, including changes in personnel, to the original
approved protocol before implementing the changes, and that significant changes will require
a new protocol.
4. If the protocol is to be discontinued, the IACUC will be notified.
5. The use of animals in this protocol cannot begin until the protocol has been reviewed and
approved by Schepens IACUC.
Principal Investigator Signature
Date
Position
Print Name:
I2. ASSURANCE OF ANIMAL FACILITY STAFF ASSISTANCE OR MONITORING.
CHECK ALL THAT APPLY:
Section C2
Section D1g
Section D4
Approval signature of Animal Facility Associate
Director:
Date:
I3. ASSURANCES OF HEALTH AND SAFETY REGISTRATION OF BIOHAZARDOUS
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MATERIAL
The Health and Safety Coordinator assures the IACUC and the animal facility staff that the use of
biohazardous materials in this study and listed in Appendix A will be handled in accordance with
applicable NIH policies, OSHA standards, Federal, state, and local regulations; and the information
provided about COMS registration numbers, animal biosafety levels, practices and procedures required
for the safe handling and disposal of contaminated animals and materials are correct.
Approval signature of the Health and Safety
Coordinator:
Date:
ASSURANCES OF HEALTH AND SAFETY REGISTRATION OF CHEMICAL HAZARDS
The Health and Safety Coordinator assures the IACUC and the animal facility staff that the use of
chemical hazardous agents in this study and listed in Appendix A will be handled in accordance with
applicable OSHA standards, Federal, state, and local regulations.
Approval signature of the Health and Safety
Coordinator:
Date:
ASSURANCES OF HEALTH AND SAFETY REGISTRATION OF RADIOACTIVE
MATERIALS USED IN LIVE AIMALS.
The Health and Safety Coordinator assures the IACUC and the animal facility staff that the use of
radioactive active materials used in live animals and listed in Appendix A will be handled in accordance
with applicable Nuclear Regulatory Commission (NRC) policies, Federal, state, and local regulations.
Approval signature of the Health and Safety
Coordinator:
Date:
ASSURANCES OF HEALTH AND SAFETY REGISTRATION OF PHYSICAL HAZARDS (i.e.,
Lasers)
The Health and Safety Coordinator assures the IACUC and the animal facility staff that the use of lasers
in live animals and listed in Appendix A will be handled in accordance with applicable Federal, State, and
Local regulations.
Approval signature of the Health and Safety
Coordinator:
Date:
Revised 6/18/2013
PI:
Protocol Number:
Appendix A: Hazardous Materials and Lasers
If you are using hazardous materials in live animals, please provide answers to the applicable questions
below:
(1) Please attach all relevant SASPs (Special Animal Safety Protocol)
(2) Radioactive materials used in live animals. Radiation Safety Committee meets quarterly.
Radiation Safety Committee review date:
(3) Biological materials used in live animals. Provide copies of the approved registration
documents from COMS, if applicable, and attach to the animal protocol.
COMS #:
(Biologicals include, but are not limited to: human infectious agents, recombinant DNA, exempt select
agents, biological toxins, and the use of human or nonhuman primate tissues or cell lines in animals.
Zoonotics are not registered with COMS.)
** Effective January 19, 2011 the NIH revised their Guidelines involving the breeding of transgenic
rodents housed under BL1 conditions. This type of breeding experiment (with two exceptions – see
below) is now considered EXEMPT under the NIH Guidelines and no longer must be registered with the
IBC (COMS). The two types of breeding experiments that must still be registered with and approved by
the IBC (COMS) under SECTION III-E of the NIH Guidelines are:

Those breeding experiments involving transgenic rodents that contain more than 50% of the genome
of an exogenous eukaryotic virus from a single family, and

Those breeding experiments in which the transgene is under the control of a gammaretroviral long
terminal repeat (LTR). (Please see SERI Safety Officer, Kathleen Gallagher with any questions)
(4) Hazardous chemical agents including, but not limited to, mutagens, teratogens and carcinogens.
Differentiate use in
the lab from use in the animal facility.
(5) Lasers used in live animals.
NOTE: Class 3B and Class 4 laser users must have Safety training thru Harvard before using the
laser and complete the SERI Laser Safety Assurance Form (please see SERI Safety Officer, Kathleen
Gallagher)
(6) For each hazardous agent, list the agent, dose, route of administrations and frequency of
administration. Please put only one agent per line.
Agent
Dose/Kg body
weight
Volume
Route of
Administration
Frequency and
Duration of
Administration
The Policy for Non-Pharmaceutical Grade Compound Use can be found at:
http://squeek.eri.harvard.edu/iacuc.html
Pharmaceutical
Grade*
If ‘No’, please fill
out Appendix C
N/A
N/A
N/A
N/A
N/A
N/A
Appendix A: Hazardous Materials (continued)
(7) Please list the potential health risks to humans and/or animals for each of the agents listed
above? Be specific.
(8) Describe the practices and procedures required for the safe handling and disposal of
contaminated animals and materials associated with this study. For radionucleotide use,
describe the methods for monitoring radioactivity and for radioactive waste removal.
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Protocol Number:
Appendix B: Surgical Procedures
Please provide a separate Surgical Appendix for EACH surgical procedure. The appendix below
can be copy and pasted as many times as needed.
Principal Investigator:
Procedure(s) to be performed:
Multiple surgeries performed on the same animal?
Yes
If yes, please indicate the time frame from one surgery to the next:
No
Please describe criteria to ensure animals have recovered from the 1st surgery before the next
takes place:
Person(s) responsible for performing surgeries:
Phone number:
Emergency phone number:
Species:
Anesthetic(s) to be used:
Dose:
Topical anesthetic to be used:
Duration:
Analgesics to be used:
Duration:
Dose:
Dose:
Route of Administration:
Route of Administration:
Route of Administration:
Frequency:
Frequency:
Frequency:
Aseptic technique: Please check the boxes to indicate which aseptic techniques will be used in this
protocol:
Animals will be clipped around the area of incision
Site will be scrubbed with betadine and wiped with alcohol
Sterile gloves will be used
Instruments will be sterilized via:
Autoclave
Chemical Please specify:
Please note that alcohol is not a sterilant
Other: Please specify:
Description of surgical procedure(s):
Appendix B: Surgical Procedures (continued)
Description of post-operative care:
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APPENDIX C: USE OF NON-PHARMACEUTICAL GRADE AGENTS
The Policy for Non-Pharmaceutical Grade Compound Use can be found at:
http://squeek.eri.harvard.edu/iacuc.html
A pharmaceutical grade compound is defined by the USDA and OLAW as an agent, which is approved
by the FDA or for which a chemical purity standard has been established by the United States
Pharmacopeia-National Formulary (USP/NF), or British Pharmacopeia (BP).
If a non-pharmaceutical grade (NPG) agent is to be used, its use must be justified as well as assurance of
its safe use in animals.
Please answer the following questions for each agent. The following information can be copy and pasted
as many times as need.
1) Name of agent or diluent:
2) Is a pharmaceutical grade agent available:
Yes
No
If yes, please justify why a NPG agent is to be used (Note: cost cannot be the determining factor)
3) Please indicate that the highest-grade agent will be utilized:
4) If known, please indicate the stability, that the agent will be pH neutral upon administration as well
as any miscellaneous information such as pyrogenicity to ensure the agent will be safe upon
administration:
5) Please describe how the lab will ensure that the agent is sterile upon administration (autoclaved, filter
sterilized, etc.):
6) Reference(s) to ensure that this agent is safe to use as formulated and that the dose is correct for this
species (Note: prior experience is also an acceptable answer):
Revised 6/18/2013
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Protocol Number:
APPENDIX D: RECOMMENDED ANESTHETIC, AND TRANQUILIZER AGENTS
For Mice:
Type of Drug
Anesthetic
Anesthetic
Tranquilizer
Dose (mg/kg-bodyweight)
Route of
Administra
tion
IP
Ketamine HCl/Xylazine HCl
100 to 200 mg/kg Ketamine combined with 20 mg/kg
Xylazine
Isoflurane
Vaporized using a nose cone, or 2- 4% using a precision
vaporizer delivered in 100% O2
Acepromazine
2-5 mg/kg
IH, to effect
IP
For Rats:
Type of Drug
Dose (mg/kg-bodyweight)
Anesthetic
Ketamine HCl/Xylazine HCl
40 to 80 mg/kg Ketamine combined with
10 mg/kg Xylazine
Isoflurane
Vaporized using a nose cone, or 2- 4% using a
precision vaporizer delivered in 100% O2
Acepromazine
1-2 mg/kg
Anesthetic
Tranquilizer
Route of
Administration
IP
IH, to effect
IP
For Rabbits:
Type of Drug
Dose (mg/kg-bodyweight)
Anesthetic
Ketamine HCl/Xylazine HCl
30-50 mg/kg Ketamie combined with 5-10
mg/kg Xylazine
Isoflurane
2-4% using a precision vaporizer delivered in
100% O2
Acepromazine
1-2mg/kg
0.1-0.75 mg/kg if given in conjunction with
ket/xyl
Anesthetic
Tranquilizer
Route of
Administration
IM
IH, to effect
IM
APPENDIX E: RECOMMENDED ANALGESIC AGENTS
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PI:
Protocol Number:
Drug Name
Type
Dose
Route of
Administration
Duration
Other Info
Opioid
0.05-0.1mg/kg
SC
8-12 hours
Give 0.1ml of the
1:10 diluted of
stock *per 30gm
mouse
Proparacaine 0.5%
Ophthalmic
Solution
Meloxicam
Topical
Anesthetic
1-2 drops to
effect
NSAID
5-10mg/kg
Apply to the cornea
surface prior to starting
eye procedures
SC
Re-apply every
15 minutes until
procedure ends
24 hours
Bupivicaine 0.25%
Local
Anesthetic
~2mg/kg
Local injection prior to
surgery-inject around
incision, infusion into
tissue
12 hours
Buprenorphine
Opioid
0.05-0.1mg/kg
SC
8-12 hours
Ketoprofen
NSAID
5mg/kg
SC
24 hours
Proparacaine 0.5%
Ophthalmic
Solution
Carprofen
Topical
Anesthetic
1-2 drops to
effect
NSAID
5mg/kg
Apply to the cornea
surface prior to starting
eye procedures
SC
Re-apply every
15 minutes until
procedure ends
12 hours
Meloxicam
NSAID
1mg/kg
SC
24 hours
Bupivicaine 0.25%
Local
Anesthetic
~2mg/kg
Local injection prior to
surgery-inject around
incision, infusion into
tissue
12 hours
Buprenorphine
Opioid
0.01-0.5mg/kg
SC
8-12 hours
Proparacaine 0.5%
Ophthalmic
Solution
Carprofen
Topical
Anesthetic
1-2 drops to
effect
NSAID
1.5mg/kg
Apply to the cornea
surface prior to starting
eye procedures
PO
Re-apply every
15 minutes until
procedure ends
12 hours
Ketoprofen
NSAID
3mg/kg
SC
12 hours
Mice
Buprenorphine
Dilute 50% with
sterile injectable
saline. Generic
name: Maracaine
Rats
Dilute 50% with
sterile injectable
saline. Generic
name: Maracaine
Rabbits
*Use sterile 5% dextrose (D5W) only for filution. Diluted Buprenorphine is good for 2
months and then must be discarded.
Revised 6/18/2013
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APPENDIX F: RECOMMENDED OPHTHALMICS, LUBRICANTS AND
ANTIBIOTICS
The list below includes additional pharmaceutical grade compounds that are available through the
Animal Facility. The Animal Facility should be contacted regarding questions on the availability of
pharmaceutical grade compounds.















Proparacaine (0.5%)
Atropine (1%)
Phenylephrine
Tropicamide (1%)
Cyclopentolate (1%)
Timolol (0.25%)
GONAK (2%)
Genteal Gel
AK-SPORE (Triple Antibiotic Ointment)
Artificial Tears
Ofloxacin
Baytril
Ditrim
Sterile Saline (9%)
Sterile Dextrose (5%)
APPENDIX G: AN EXAMPLE JUSTIFYING THE NUMBER OF
ANIMALS NEEDED (SECTION C1)
Revised 6/18/2013
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Experimental Set 1:
Is the NALP3 inflammasome required for development of CNV following lipid injection?
Rationale: The innate immune system utilizes a variety of receptors that can recognize host-derived
danger signals that are released from a sick or injured cell. NOD-like receptors (NLRs) are one of these
receptors and are critical in activating inflammation. Activation of the NLRs leads to the formation of a
multiprotein complex known as the inflammasome that upon activation produces the proinflammatory
cytokine IL-1beta. Both NALP3 and ASC are critical components of the inflammasome and we will use
mice that have either the NALP3 or ASC components knocked-out to determine the function of the
inflammasome in the development of CNV using a mouse model of AMD. The Mann-Whitney U test of
proportions will be used to find statistical relevance. This requires no less than 10 total mice per
treatment group (5 mice per group repeated 2 times). The groups of mice are as follows:
Experimental groups:
C57BL/6 Wild-type (controls)
C57BL/6-NALP3-KO
C57BL/6-ASC-KO
Mouse numbers: 3 groups of mice x 2 (lipid pr borate buffer alone) x 6 time points (-1, day 3, 1 week, 2
weeks, 3 weeks, 5 weeks) x 2 (retinal whole mounts and immunohsitochemistry on frozen sections) x 5
per group x 2 (repeat once for reproducibility) = 720 mice
Experimental Set 2:
Is membrane FasL a critical mediator of CNV in the pathogenesis of Wet AMD?
Rationale: Fas ligand is an important protein in maintaining immune privilege in the eye by preventing
inflammation. Fas ligand exists as a membrane-bound protein and a soluble protein that is released from
the cell surface. Our laboratory has shown that membrane FasL and soluble FasL have opposing
functions; membrane FasL is pro-apoptotic and pro-inflammatory, while soluble FasL is anti-apoptotic
and anti-inflammatory. However, it is unclear how the different forms of FasL contribute to the
development of CNV. To address this question, we will use mutant mice that do not express any Fas
ligand (Fas ligand knock-out) and mice that only express the membrane form of FasL and do not express
soluble FasL.
Experimental groups:
C57BL/6 Wild-type (we will use the data from controls above)
C57BL/6-membrane FasL only (DCS)
Balb/c-Wild-type (controls)
Balb/c-FasL-KO
Mouse numbers: 3 groups of mice x 2 (lipid pr borate buffer alone) x 6 time points (-1, day 3, 1 week, 2
weeks, 3 weeks, 5 weeks) x 2 (retinal whole mounts and immunohsitochemistry on frozen sections) x 5
per group x 2 (repeat once for reproducibility) = 720 mice
Breeding:
To perform these studies we will need approximately 80 mice/year from each strain. The FasL knockout
(homozygous) and mFasL-only (homozygous) mice breed well and produce about 6 pups per litter and
each female has about 4-5 litters per year. Therefore, to ensure we will get 80 mice/year, we will set up 3
breeder cages (2F:1M). These breeder cages will be replaced every 6 months. Therefore, for breeding we
will need 54 mice per strain: 3 breeder cages x 3 mice per breeder cage x cages replaced 2 times/year x 3
Revised 6/18/2013
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Protocol Number:
years = 54 mice
54 homozygous FasL knockout mice for breeding
54 homozygous membrane FasL-only mice for breeding
breeding total: 108
Total mice:
C57BL/6 wild-type:
C57BL/6-NALP-3 KO:
C57BL/6-ASC-KO:
Membrane FasL only:
Breeders:
Balb/c wild-type
Balb/c-Fasl-KO:
Breeders:
Total:
Revised 6/18/2013
240
240
240
240
54
240
240
54
1548
PI:
Protocol Number:
APPENDIX H: AN EXAMPLE FOR EXPERIMENTAL TIMELINE AND
PROCEDURE NARRATIVE (SECTION D)
Experimental design:
(Day 0) Anesthetize mice and inject HpODE lipid or Borate buffer (Vehicle control) subretinal
(Day -1, Day 3, Week 1, Week 2, Week 3, and Week 5 post injection) Anesthetize mice and perform
OCT followed by Fluorescein fundus angiography
groups of mice are euthanized and eyes
enucleated at each time point for retinal whole mounts and immunohistochemistry on frozen sections.
*** because the NALP3-KO and ASC-KO mice will be housed in the SERI satellite facility (see special
care section D5), the OCT and fluorescein studies on the NALP3-KO and ASC-KO mice will be
performed at the MEEI facility. All other mice will be examined using the SERI facility.
Subretinal injection of LipidMice will be deeply anesthetized with an intraperitoneal injection of ketamine (120 mg/kg) and xylazine
(20 mg/kg). The pupils will be dilated with topical 1% tropicamide to view the fundus, and the eye treated
with topical anesthetic (proparicaine drops). Subretinal injections are made under transpupillary
observation using a binocular surgical microscope. The conjunctiva adjacent to the cornea is grasped
with forceps to allow optimal exposure of the injection site and stabilize the eye. A 30-guage needle is
used to penetrate the sclera approximately 2 mm posterior to the limbus, while being cautious not to
damage the lens during the procedure. A retinotomy (retinal incision) is performed in the peripheral
retina with the tip of the subretinal injector (Glaser subretinal injector (20-gauge shaft with a 32-gauge
tip) and 2ul of HpODE (30ug in borate buffer Ph9) or borate buffer alone (vehicle control) is slowly
injected into the subretinal space. Once the injection is complete, a bacitracin ophthalmic topical ointment
will be applied to the injection site in order to guard against infection
Optical Coherence Tomography (OCT)Mice will be deeply anesthetized with an intraperitoneal injection of ketamine/xylazine (120 mg/kg
Ketamine combined with 20 mg/kg Xylazine). The pupils will be dilated with topical 1% tropicamide to
view the fundus, and the eye treated with topical anesthetic (proparicaine drops). Puralube ointment will
be placed on the contralateral eye to prevent any drying of the cornea. After anesthetization, the mouse
will be restrained in a mounting tube that is fixed on a six-axis platform. The fundus camera in the optical
head of the apparatus will provide initial alignment for the sample light, to ensure it is delivered through
the dilated pupil. Final alignment will be guided by monitoring and optimizing the real time OCT image
of the retina, with the whole set up procedure taking approximately 5 minutes for each mouse eye. A
complete scan only takes a few seconds and it is a non-contact procedure.
Fluorescein Fundus AngiographyImmediately following the OCT exam, a drop of sterile saline will be placed on the experimental eye to
remove any debris followed by a contact lens. Then 0.01ml of 25% sodium fluorescein (pharmaceutical
grade sodium fluorescein; Akorn Inc)/5 g body weight is injected i.p. The retinal vasculature will fill with
dye in less than one minute following injection. Photos will be taken sequentially at 1, 2, 3, 4 and 5
minutes post fluorescein injection.
****No genotyping is required b/c all mice are homozygous
Revised 6/18/2013
PI:
Protocol Number:
APPENDIX I: FLOW CHART EXAMPLES (SECTION C)
Description of Experimental Groups and Controls
Experiment set #3
88 mice
44mice
44 mice
Wild-type (PGC1a
+/+
)
Laser-induced CNV, OCT
OCT post-CNV and
fluorescein angiography
Day 7
Laser-capture
microdissection
5 mice, day 7
IF and
morphology
5 mice, day 7
12 mice, day 7
5 mice, day 14
IF and
morphology
5 mice, day 14
Whole retina
protein and RNA
12 mice, day 14
Revised 6/18/2013
Laser-induced CNV, OCT
OCT post-CNV and
fluorescein angiography
Day 7
Whole retina
protein and RNA
Laser-capture
microdissection
-/-
PGC1 alpha null (PGC1a )
Laser-capture
microdissection
5 mice, day 7
IF and
morphology
5 mice, day 7
Whole retina
protein and RNA
12 mice, day 7
Laser-capture
microdissection
5 mice, day 14
IF and
morphology
5 mice, day 14
Whole retina
protein and RNA
12 mice, day 14
PI:
Protocol Number:
APPENDIX J: EXPERIMENTAL TIME LINE EXAMPLE (SECTION D)
Anesthetize and apply analgesia  perform corneal epithelial debridement on right eye 
apply adequate post-operative care (antibiotic ointment and Buprenex)  monitor wound
healing for determined time frame (maximum of 96 hours)  anesthetize (4, 24, 48, 72, or 96
hrs)  stain cornea with fluorescein  observe with indicated instruments for analysis 
euthanize  enucleate manipulated eye  perform morphologic evaluation of the tissue or
RNA isolation.
No methods of restraint other than hand held will be used. Animals will be identified using an
ear punch, a method approved by NIH Office of Laboratory and Animal Welfare.
Revised 6/18/2013
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Protocol Number:
APPENDIX K: AN EXAMPLE FOR SEARCHES FOR ALTERNATIVES
TO PROCEDURES THAT MAY CAUSE PAIN AND DISTRESS (SECTION F)
There are two procedures that require searches for alternatives:
1. Immunization with Freund’s adjuvant (potential pain and distress).
1.
2.
3.
4.
5.
Procedure: Freund’s adjuvant
Keywords Freund’s adjuvant Th1 cells, mice
Database searched Pubmed
Date of the search: 10/7/08
Period of time covered by the search: 1950 – October, 2008
6. The search resulted in no references. Moreover, we provided an abstract of a report from a group of
NIH researchers describing the requirement of Freund’s adjuvant in an antigen immunization to
generate a Th1 cell response. To immunize without adjuvant promotes immunity that does not induce
the T cell response. To immunize without adjuvant promotes immunity that does not induce the T
cells responsible for autoimmune disease, graft rejection, and hypersensitivity. These T cells are
called Th1 cells, and adjuvant is called Freund’s (complete) adjuvant. We have attached the search
results.
For help with your literature search or additional databases contact the Animal Welfare Information
Center at: www.nal.usda.gov/awic/databases/database.htm or (301) 504-6212 or awic@nal.usda.gov.
2. Experimental Autoimmune Uveoretinitis (EAU), (potential distress)
1. Procedure: Experimental autoimmune uveoretinitis
2. Keywords; Uveitis, mouse
3. Database searched: Association for Research in Vision and Ophthalmology (ARVO) and American
Association of Immunologist (AAI)
4. Date of the search: May 4 and 6, 2003
5. Period of time covered by the search: 2003
6. A search of abstracts for mouse models of uveitis presented at the annual meetings of the American
Association of Immunologist (AAI; May 6, 2003) and the Association for Research in Vision and
Ophthalmology (ARVO; May 4, 2003) showed no alternative models for experimental autoimmune
uveitis other than the procedures described in this protocol. The incidence of uveitis in both eyes is
unavoidable, however, our work involves a possible treatment that reduces the incidence of the
disease and the severity of inflammation in eyes of mice made to express uveitis. Since mice mainly
use smell to guide themselves to food, we have not observed uveitic mice failing to feed themselves.
Revised 6/18/2013
PI:
Protocol Number: