August 5, 2016

Manual for biological experiments (second edition)—For SPring-8/SACLA users

Safety Office, Japan Synchrotron Radiation Research Institute (JASRI)
Harima Safety Center, RIKEN

August 5, 2016

Contents
1. Introduction－－－－－－－－－－－－－－－－－－－－－－－1

2. Precautions for handling biomaterials －－－－－－－－－－－－1

3. Experiments involving microorganisms－－－－－－－－－－－－3

4. Genetic recombinant experiments－－－－－－－－－－－－－－7

5. Animal experiments －－－－－－－－－－－－－－－－－－－11

6. Research involving human subjects－－－－－－－－－－－－－16

7. Measures to be taken in case of accidents －－－－－－－－－－19

August 5, 2016

1. Introduction
This textbook provides, in order to ensure the safety of research conducted at SPring-8/SACLA,
health and safety knowledge and the rules for the facility users engaged in biological experiments.
In biological experiments, various biological materials are used. Before drafting a
research/experiment plan, you must understand all the properties of the biological material you aregoing to use and learn how to use the devices and the like prior to starting an experiment.
Biological experiments, including genetic recombinant experiments, animal experiments,
experiments involving microorganisms and research involving human subjects (human derived
samples/information), must conform to the laws, regulations, guidelines, and the rules specified bySPring-8/SACLA. Please note that you must observe these rules to ensure the safety of workers
engaged in such experiments, other researchers and the general community.
Research involving human subjects (safety matters conform to those of microorganism
experiments) and animal experiments should be conducted with full consideration given to research
ethics and animal welfare respectively.

2. Precautions for handling biomaterials
To conduct research involving biomaterials in a safe manner, observe the following two major
points:(1) Consider risks of damage to the environment, including beyond the location at which theexperiment is performed, and risks to living organisms, including workers engaged inexperiments, caused by microorganisms and biologically active agents contained inexperimental biomaterials.
(2) Use experimental equipment and tools in a proper manner.

2-1. Infection, allergies and poisoning due to biological materials(1) Infection by biomaterials
Handling microorganisms and experimental animals involves the risk of infection. Always wear
appropriate protective gear such as a laboratory coat, rubber gloves, protective glasses and a mask
to prevent infection. In order to avoid secondary infection, inactivate fungus bodies and culture
fluid using an autoclave and the like before disposal.

(2) Allergies due to biomaterials
When handling materials that contain allergens, use mechanical pipettes and wear protective gear
including a laboratory coat, rubber gloves and protective glasses to avoid direct contact with thematerials. Waste that contains allergens must be put in a sealed plastic bag and the like to bedisposed of safely.

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Handle experimental animals wearing appropriate protective gear because experimental animal
hair, dust from the breeding rooms and animal bites can be allergens.

(3) Poisoning caused by biomaterials
When conducting biological experiments using toxins and the like produced by organisms,
handle them, in a similar way to handling dangerous chemicals, carefully and while wearing
appropriate protective gear, including rubber gloves.

2-2. Handling biochemical reagentsSome reagents used in biological experiments possess high carcinogenicity, teratogenicity,
neurotoxicity, and allergenicity. Wear proper protective gear when using such reagents.
After processing biomaterials containing such substances in an autoclave, wait for them to cool
down before removing them, as vapor from items that are taken out of the device shortly after
autoclaving may contain inactivated substances.
When handling poisonous and deleterious substances, refer to the following websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/chemistry (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/chemistry (in English)

2-3. Handling experimental/analytical devices
In order to use devices in a safe manner, refer to the manuals and, before handling complex devices
in experiments, have an experienced person demonstrate how to use it. Confirm that reagents and thelike used in a device are set in place.
In biological experiments, the following accidents have been reported: i) burns caused by heating
gel with a microwave oven, ii) burns resulting from contact with a hot autoclave, iii) injury of theeyes during UV radiation while not wearing protective glasses for UV rays, and iv) fire/smoke
resulting from improper setting of the temperature of a dry heat sterilizer.
Accidents were also caused by improper handling of injection needles, surgery instruments,
microtome blades, and other tools with sharp edges. Workers must handle experimental equipment
carefully and dispose of it in a proper manner.

When handling microwave ovens, autoclaves, and transilluminators,
wear appropriate protective gear.
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Be careful to set the temperatureof dry sterilizers.

Wear cut-resistant gloves whenhandling blades.

Handling liquid nitrogen, which is often used for instant freezing of biomaterials, also causes
accidents. Examples include frostbites due to contact with liquid nitrogenand rupture of freezing tubes being thawed. When handling liquid
nitrogen, wear protective glasses and cryo-protective gloves while taking
necessary safety measures. Adequately ventilate the room where liquid
nitrogen is being used. Using liquid nitrogen in a closed place can cause
suffocation.
Experiments using the following items must conform to related laws andregulations and guidelines, and rules specified by the SPring-8/SACLA:i) dangerous devices that produce high temperature, vapor, high pressuregases, UV rays, high voltage, and laser beams, and ii) labelled compounds containing radioisotopes.
When conducting such experiments, observe these laws, rules and the like.

3. Experiments involving microorganisms
Microorganisms, etc.* should be handled in conformity with related laws, regulations, and the rules
specified by SPring-8/SACLA. Only level 1 and 2 microorganism experiments are allowed to be
conducted on the Harima Campus.

* Microorganisms, etc. refer to bacteria including chlamydia and rickettsia, viruses, viroid,3

August 5, 2016

fungi, parasites and biological materials (including human derived samples) contaminated withthem, but exclude genetically modified organisms.

Levels of microorganisms, etc.
Level 1Have no risks of causing significant infection to humans or animals
Pathogenic to humans and animals but have a low possibility of causing

Level 2

serious danger inside the laboratories, to other workers, and to theenvironment.

3-1. Required procedures
Before handling microorganisms in research in SPring-8/SACLA, the procedures shown below
must be followed:(1) Applying research plans
When using materials that can damage the human body, livestock, and agricultural and marine
products through biological interaction, submit the Biological Experiment Plan and the Safety
Instruction Manual to the Users Office of Japan Synchrotron Radiation Research Institute (JASRI)
by the designated date. Materials that require submission include: pathogenic microorganisms
such as bacteria, viruses, fungi, and infectious nucleic acids/plasmids; parasites and toxic
substances produced by them; carcinogens, allergens, and other similar materials."For details,
refer to the following websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)

When handling human derived samples, the procedures for an experiment involving humansubjects must also be followed, in addition to the procedures above. See 6-1. Required proceduresin 6. Research involving human subjects.

(2) Handling legally regulated microorganisms, etc.
Before handling i) export prohibited items specified by the Plant Protection Act, ii) specificpathogens designated by the Act on Prevention of Infectious Diseases and Medical Care forPatients Suffering Infectious Diseases, iii) pathogens specified by the Act on Domestic Animal
Infectious Diseases Control, and iv) microorganisms regulated by other laws and regulations, youmust submit an application for approval or a notification to the government authorities. When you
are planning to use such items, consult JASRI Safety Office as early as possible.

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(3) Carrying in microorganismsa) Level 1 microorganisms
Submit the List of Samples, Reagents, etc. via my page on the User Information page ofSPring-8/SACLA no later than 10 days before each visit.
https://user.spring8.or.jp/apps/login/en?institution=0 (in English)b) Level 2 microorganisms
Submit a written Transport of Infectious Agents Designated Biosafety Level 2 to JASRI Users
Office no later than two weeks prior to the date of carry-in. For details, refer to the following
websites:
SPring-8: https://user.spring8.or.jp/?p=1565 (in Japanese)
SACLA: http://sacla.xfel.jp/?p=89 (in Japanese)
SPring-8: https://user.spring8.or.jp/?p=1565&lang=en (in English)
SACLA: http://sacla.xfel.jp/?p=89&lang=en (in English)

Submit to: JASRI Users Office, tel: 0791-58-0961, email: sp8jasri@spring8.or.jp

3-2. Precautions for conducting experiments(1) Location of experiments
Microorganisms must only be handled in approved experimental rooms, where drinking, eating
and storing food are prohibited.

(2) Signs
Post an international biohazard sign and a label indicating the level of the microorganisms at theentrance to the laboratories and storage cabinets where microorganisms are actually handled andstored.

Post the signs at theentrance.

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(3) Containment measures/infection preventive measures
When conducting experiments using microorganisms, observe the following rules to prevent the
spread of contamination, damage to the environment, and infection accidents.
Note that human derived samples are classified into level 2 and must be handled as infectious
material. When handling infectious viruses such as hepatitis B viruses, you must adequately
consider i) equipment, ii) methods, iii) incubation period, and iv) the possibility that otherinfectious known and unknown viruses are present.

1) Prepare disinfectants and sterilizers appropriate for the microorganisms you use.
2) Avoid wearing clothes, sandals and the like that allow skin to be exposed.
3) Wear appropriate protective gear such as a laboratory coat, rubber gloves, protective glasses,
and a mask.
4) Close the windows and doors of the experimental room to prohibit entry of non-authorized
persons.
5) When handling microorganisms, the use of a biohazard cabinet is recommended.
6) Keep the location of experiments in order.
7) Disinfect and sterilize inside of equipment and tools, and wash your hands when necessary.
8) When taking microorganisms out of an experimental room, put them in a container thatprevents them from leaking and spreading.
9) When using microorganisms in the experimental hall (including the experimental hutch),take necessary measures to avoid scattering of them. In the experimental hutch, where theirleakage may be caused by beam radiation, take necessary measures to avoid scattering ofthem.
10) Store solid/liquid waste in a cool dark place after disinfecting/sterilizing or sealing up.

When handling level 2 microorganisms, also take the following measures:

11) Explain to others working in the vicinity all of the hazardous properties of the
microorganisms and the like that you use prior to starting an experiment.
12) When handling microorganisms, take measures to limit aerosols to the least amountpossible. When aerosols are generated or when there is such a possibility, conduct
experiments in a biohazard cabinet.

(4) Storage
Levels 1 and 2 microorganisms should be stored using dedicated storage containers in approved
experimental rooms. If such an experimental room is vacant, lock the storage cabinets or the room.

6August 5, 2016

When they are stored in a sealed container that can avoid leakage of the contents, or when othersimilar measures are taken, they can be stored in a storage cabinet outside of the experimentalroom where they are actually used.

(5) Disposal
Disinfect and sterilize waste originating from your experiment and bring it back to yourlaboratory to dispose of.
Limit the items that can cause infection to the least amount possible and disinfect/sterilize them
totally before disposal to prevent contamination and spreading.
Disinfect/sterilize used biomaterials such as bacterial cells/culture fluid and disposable
experimental tools that are stained with such materials using autoclaves and the like before
disposal.
Before washing laboratory ware, always disinfect and sterilize it. The primary and secondary
rinse water should be collected as waste liquid.

4. Genetic recombinant experiments
They must be conducted in conformity with related laws and regulations and the rules specified bySPring-8/SACLA.
On the Harima campus, only the following experiments are allowed to be conducted: i)experiments that require P1 and P2 level containment measures, ii) P1A and P2A animal
experiments and iii) P1P and P2P experiments using plants.

4-1. Required procedures
When conducting genetic recombinant experiments at SPring-8/SACLA, the following procedures
are required:(1) Applying research plans
When conducting genetic recombinant experiments at SPring-8/SACLA, apply a research planand undergo the committee’s examination process to obtain approval. Submit the Application forApproval of Genetic Recombination Experiment to JASRI Users Office by the designated date.
For details, refer to the following websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)

When handling animals (mammals, birds, and reptiles), submit the Animal Experiment
Application Form to JASRI Users Office, separately from obtaining approval of the Application
for Approval of Genetic Recombination Experiment, to have it examined by the committee for7August 5, 2016

approval as an animal experiment. See 5-1. Required procedures in 5. Animal experiments. For
more details, refer to the following websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)(2) Carrying in/out genetically modified organisms
Submit a written Genetically Modified Organisms Carry In/Out Notification Form to JASRI
Users Office at least ten days prior to the date of work. For more details, refer to the following
websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)

Submit documents to: JASRI Users Office, tel: 0791-58-0961, email: sp8jasri@spring8.or.jp

4-2. Precautions for conducting experiments(1) Location of experiments
Genetic recombinant experiments must be conducted in approved experimental rooms with thedoors closed. In order to prevent persons who are not engaged in the experiments from entering
the room without permission, post a sign indicating authorized personnel only at the entrance tothe room, lock the room when it is vacant and take other necessary security measures.
Experiments using genetically modified animals/plants must be carried out in a location that isdesigned to prevent such animals and pollen of such plants from escaping and scattering.
Drinking, eating and storing foods are prohibited in experimental rooms.

(2) Signs
For P2, P1A, P2A, P1P and P2P level experiments including culturing, breeding, and raising
genetically modified organisms, the signs shown in table 1 are required to be posted, dependingon the content of experiments.
When storing genetically modified organisms intended for use in experiments in an experimentalroom temporarily, which is legally considered as “experiment underway”, take the same measures.
The signs must be also posted at the storage facilities.
When a sign indicating experiment underway is posted, close the door of the experimental room.
If you leave the door open while experiments are not being conducted in the room, remove or
cover the signs, and take other measures to make them invisible.

Table 1 Required signs and location to post8August 5, 2016

Level

P1A

Sign

Location to post

組換え動物等飼育中

Entrance to experimental room

Genetically modified animals now breeding

P1P

組換え植物等栽培中

Entrance to experimental room

Genetically modified plants now raising

P2

P2レベル実験中

Entrance to experimental room andP2 level experiment going on

P2A

組換え動物等飼育中

storage equipment

Entrance to experimental room

Genetically modified plants now raising

P2P

組換え植物等栽培中

Entrance to experimental room

Genetically modified plants now raising

組換え動物等飼育中

Example of posting signs at the entrance to P1A experimental room
(genetic recombinant experiments)(3) Containment measures
When handling genetically modified organisms in SPring-8/SACLA, which falls under acategory of Type 2 Use under the laws, take containment measures that are specified by the laws
and regulations or approved by the Minister of Education, Culture, Sports, Science andTechnology. Please observe the following rules:

1) Prepare agents and sterilizers that can adequately inactivate the genetically modified

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organisms you use.
2) Avoid wearing clothes, sandals and the like that allow skin to be exposed.
3) Wear appropriate protective gear such as a laboratory coat, rubber gloves, protective glasses,
and a mask.
4) Close the windows and doors of experimental rooms where the experiments are being
conducted.
5) When handling genetically modified animals, take appropriate escape-prevention measuresand containment measures depending on each animal species (for example, rodent barriers).
6) When performing an operation that can generate aerosols and scatter genetically modified
organisms and the like, use tools, devices, and equipment that prevent biohazard.
7) Operations in P2 level experiments that can generate aerosols should be conducted in abiohazard safety cabinet.
8) Keep the location of experiments in order.
9) Disinfect and sterilize inside of the equipment and tools, and wash your hands whennecessary.
10) When equipment (including a lab bench), devices, and tools are contaminated withgenetically modified organisms, or when there is such a possibility, disinfect and sterilize
these items after experiments.
11) When transferring genetically modified organisms, put them in a container that can prevent
them from leaking, escaping, and scattering.
12) When using genetically modified organisms in the experimental hall (including theexperimental hutch), take necessary measures to avoid scattering of them. In theexperimental hutch, where their leakage may be caused by beam radiation, take necessary
measures to avoid scattering of them.
13) Store solid/liquid waste in a cool dark place after disinfecting/sterilizing or sealing up.

(4) Storage during experiments
Genetically modified organisms must be stored in an approved experimental room using
containers that can prevent them from leaking, escaping and scattering. Post a sign indicating thatP2 experiments are underway at the storage equipment where P2 level genetically modified
organisms are actually stored.

(5) Disposal
Disinfect and sterilize waste and the like originating from your experiment and bring it back toyour laboratory to dispose of.
Before disposing of genetically modified organisms, biomaterial before genetic recombinant, and10August 5, 2016

tools contaminated with such material, always disinfect and sterilize them by autoclaving and thelike.
Before washing laboratory ware, always disinfect and sterilize it. The primary and secondary
rinse water should be collected as waste liquid.

5. Animal experiments
They must conform to the related laws and regulations and the rules specified by SPring-8/SACLA.
Before conducting animal experiments, serious consideration must be given to i) purpose andnecessity of animal use, ii) prevention of unnecessary repetition, iii) search for alternative methods,iv) limiting animals to use to the least number possible, v) pain intensity felt by experimental
animals and appropriate methods to alleviate their pain. You must also improve your experimental
techniques and give full consideration to animal welfare based on 3R principles (Replacement,
Reduction, Refinement).

5-1. Required Procedures
When conducting animal experiments at SPring-8/SACLA, the following procedures are required:(1) Applying research plans
When conducting animal experiments at SPring-8/SACLA, apply a research plan and undergo
the committee’s examination process to obtain approval. Submit the Animal Experiment
Application Form to JASRI Users Office by the designated date. For details, refer to the following
websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)

When handling genetically modified animals and inoculating animals with genetically modified
organisms in experiments, in addition to obtaining approval with the Animal Experiment
Application Form, submit the Application for Approval of Genetic Recombination Experiment to
JASRI Users Office and undergo the committee’s examination to obtain approval as a genetic
recombinant experiment. When conducting infection experiments using microorganisms that are
not genetically modified, similarly, submit the Biological Experiment Plan and the Safety
Instruction Manual for an experiment involving microorganisms as well as the Animal Experiment
Application Form for an animal experiment to JASRI Users Office and have them examined by
the committee to obtain approval. Refer to 4-1. Required procedures of 4. Genetic Recombinant
experiments and 3-1. Required procedures of 3. Experiments involving microorganisms. For
details, refer to the following websites:

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http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)

Before handling narcotics, stimulants, and psychotropics, you should apply for licenses or
identification certificates to the government authorities. When you are going to handle any of such
substances, consult JSARI Safety Office as early as possible. For details, see the following
websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/chemistry (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/chemistry (in English)(2) Carrying in experimental animals
When delivering experimental animals from outside to SPring-8, submit a “notification ofdelivery” at least one week before the delivery date.
For details, refer to the following websites:
http://www.spring8.or.jp/ext/ja/bio_formmail/sfm_form_en.html (in English)
When importing experimental animals, before bringing them into SPring-8/SACLA, theprescribed procedures are required by the animal import/export notification system, which
conforms to the Act on Prevention of Infectious Diseases and Medical Care for Patients Suffering
Infectious Diseases. When you are planning such action, contact JASRI Safety Office as early as
possible.

Submit Documents to:JASRI Users Office, tel: 0791-58-0961, e-mail：uoffice@spring8.or.jp

5-2. Precautions for conducting experiments(1) Location of experiments
Use of experimental animals, that includes experiments, storage, and breeding, must be
conducted in approved experimental places. Never eat, drink, and store foods in such places.

(2) Signs
Some animal species such as macaques need legally required signs to be posted.

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特 定 動 物この動物は人の生命、身体及び財産に害を加えるおそれがある動物であるため、第三者の接触等を禁止します。

許 可 年 月 日

平成１９年５月１６日

有効期間の末日

平成２４年５月１５日

許 可 番 号

第２－３４号

特定動物の種類

ニホンザル

Sign indicating specific animals (dangerous animals) now being bredand prohibiting entry of non-authorized persons(3) Escape/infection/allergy-prevention measures
When handling experimental animals, observe the following rules:
1) Prepare disinfectants and sterilizers appropriate for the experimental animals you use.
2) Conduct an inspection to check if experimental animals are infected with pathogenic
microorganisms when necessary.
3) Monitor your health condition every day and avoid contact with experimental animals when
you do not feel well.
4) Avoid wearing clothes, sandals and the like that allow skin to be exposed.
5) Wear appropriate protective gear such as a laboratory coat, rubber gloves, protective glasses,
and a mask.
6) In order to prevent experimental animals from escaping and scattering into the naturalenvironment, take appropriate preventive measures such as placing rodent barriers, closing
the doors of experimental rooms and preparing capture tools.

Rodent barrier placed at the door

Capture net7) Use local exhaust ventilations such as biohazard cabinets when necessary.
8) Keep the location of experiments in order.
9) Disinfect and sterilize inside of the equipment and tools, and wash your hands when13

August 5, 2016

necessary.
10) Transfer experimental animals using transport containers appropriate for animal species.
(for example: put animals in a carry bag as well as their cage, use a cardboard box with wire
mesh or transport container used by experimental animal distributors.)

Transport box for

Cardboard box for transporting

experimental animals

experimental animals

11) When handling experimental animals in the experimental hall (including the experimental
hutch), take necessary measures to avoid animal escape.
12) Store solid/liquid waste in a cool dark place after disinfecting/sterilizing or sealing up.

(4) Humane endpoints and euthanasia
In experiments involving a high degree of pain, anesthesia or analgesia must be administered tothe animals. When such procedures obstruct the experiments and cannot be used however, set ahumane endpoint that can quickly relieve the animals of their pain and achieve the purpose ofexperiments.
When euthanizing experimental animals, administer an anesthetic to them exceeding its fatal
dose or take other appropriate measures to make them unconscious quickly leading to death withthe least pain possible. Make sure that the animals are dead.

(5) Experimental operations
In order to ensure animal welfare and the safety of workers engaged in animal experiments,
perform proper experimental operations and acclimate experimental animals to a newenvironment prior to handling them in experiments.
When necessary, anesthetize and restrain them in a proper manner. Effects obtained by anesthesia
and restraint depend on the experience of those who actually perform such procedures. Beginners
must receive instructions from experienced persons.
When handling narcotics, stimulants, and psychotropics, refer to the website below:

14August 5, 2016

http://www.spring8.or.jp/ja/users/safety/form_procedure/chemistry (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/chemistry (in English)

Perform the following operations with full consideration given to the matters below:

1) Anesthesia and sedation
Experiment workers should do their best to spare experimental animals’ unnecessary pain.
Use anesthetics, sedatives or tranquilizers in a proper manner and within the extent necessary
for the purpose of the animal experiment.

2) Restraint
When necessary, experimental animals should be restrained in order to relieve the pain they
may suffer, to facilitate experimental procedures or to prevent hazards for investigators. But
animals should not be restrained any longer than the minimum amount of time necessary.
Animals should be acclimated in advance to restraint devices or behavior observation devices.
Prior to restraining an animal, monitor it carefully. If it is agitated, wait until it calms down.
3) Surgery and post-surgical monitoring
Animals should be operated under anesthesia and asepsis; anesthetic monitoring should bemaintained during surgery. After surgery, anesthetic recovery and body temperature should bemonitored. Prevention of post-surgical infections must be given a high priority. When the
animal appears to experience severe pain, appropriate measures should be taken such asadministration of a sedative. If necessary, euthanasia (humane endpoint) should be considered.

(6) Monitoring experimental animals
Experiment workers should observe each experimental animal, everyday if possible, from its
arrival in the animal facility until the end of the experiment, and give adequate treatment whennecessary.
When they spot a dead animal or an animal that is experiencing a health problem, they should
notify the principal investigator as well as the animal care manager. Ailing or injured animals
should immediately receive adequate treatment or be euthanized.

(7) Procedures at the end of an experiment
When carrying out an experimental animal after an experiment, make sure that the transport
container size and the method and time of transferring are appropriate from humane and hygiene
perspectives. When the experimental animal cannot be used in another experiment, it should be
euthanized.

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August 5, 2016(8) Disposal of dead animals and the like
Carcasses and the like (including tissues) of experimental animals as well as unsanitary items
such as bedding should be appropriately stocked or disposed of, to prevent their impact on theenvironment and public hygiene.
Carcasses and the like of experimental animals must be kept frozen so that they do not putrefy
before their final disposal. Unsanitary items like used bedding should also be stocked
appropriately, to prevent their smell and the like from affecting human health and theenvironment.
Bring back carcasses of experimental animals and unsanitary items such as used bedding to yourlaboratory to dispose of after disinfecting or sterilizing them when necessary.
Before washing laboratory ware, always disinfect or sterilize it and collect the primary andsecondary rinse water as waste liquid.

(9) Care of experimental animals
Experimental animals should be taken care of lawfully and appropriately in compliance with theStandards Relating to the Care and Management of Laboratory Animals and Relief of Pain (Notice
No.88 of the Ministry of the Environment).

6. Research involving human subjects
Such research must conform to the related laws and regulations and the rules specified bySPring-8/SACLA.
When conducting the research, make sure that you i) respect the welfare of the donors, ii) respect
the intention of the donors to decide whether or not they participate in the research, iii) obtain theinformed consent, and iv) observe other ethical codes specified by the Ethical Guidelines for
Medical and Health Research Involving Human Subjects (Declaration of Helsinki adopted by theGeneral Assembly of the World Medical Association in 1964).

6-1. Required procedures
When conducting experiments involving human derived samples/information at SPring-8/SACLA,
the following procedures are required:(1) Applying research plans
When conducting an experiment involving human derived samples/information at
SPring-8/SACLA, submit the Application Form for the Handling of Human Materials to JASRI
Safety Office, no later than two weeks prior to carrying in the items, to obtain approval from the16August 5, 2016

Safety Office. For details, refer to the following websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)

Make sure that the experiment has already been approved by the institution with which you areaffiliated.
When handling human derived samples that are not identified to be non-infectious, submit the
Biological Experiment Plan Safety and the Instruction Manual to JASRI Users Office, separatelyfrom obtaining approval of the Application Form for the Handling of Human Materials, andundergo the committee’s examination to obtain approval. See 3-1. Required procedures of 3.
Experiments involving microorganisms. For details, refer to the following websites:
http://www.spring8.or.jp/ja/users/safety/form_procedure/biology (in Japanese)
http://www.spring8.or.jp/en/users/safety/form_procedure/biology (in English)(2) Carrying in/out human derived samples
Submit a List of Samples, Reagents, etc. via my page on the User Information page ofSPring-8/SACLA no later than 10 days before each visit.
https://user.spring8.or.jp/apps/login/en?institution=0 (in English)

Submit documents to: JASRI Users Office, tel: 0791-58-0961, e-mail: uoffice@spring8.or.jp

6-2. Precautions for conducting experiments(1) Location of experiments, signs, and infection prevention measures
1) Non-infectious human derived samples
They have no requirements for the location of experiments and do not need signs or infection
prevention measures.

2) Human derived samples that are not identified to be non-infectious
Using such samples requires the procedures for research involving human subjects as well as
those for research involving microorganisms.

a) Location of experiments
Human derived samples that are not identified to be non-infectious must be handled inapproved experimental places. Never drink, eat, and store foods in such places.

b) Signs

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Post an international biohazard sign and a label indicating level 2 microorganisms at theentrance to the laboratory and on the storage cabinets.

c) Infection prevention measures
Take the following measures to prevent pollution from spreading to the environment and to
avoid infection accidents:
1) Prepare appropriate disinfectants and sterilizers.
2) Avoid wearing clothes, sandals and the like that allow skin to be exposed.
3) Wear appropriate protective gear such as a laboratory coat, rubber gloves, protective
glasses, and a mask.
4) Close the windows and doors of the experimental room and prohibit entry of
non-authorized persons.
5) When there are other investigators working nearby, explain to them about the risks of thehuman derived samples you will be using in advance.
6) When handling human derived samples, take measures to limit aerosols produced in theexperiment to the least amount possible. Experiments that can generate aerosols must be
conducted in a biohazard cabinet.
7) Clean and organize the experimental rooms.
8) Disinfect and sterilize inside of the equipment and tools, and wash your hands whennecessary9) When bringing out human derived samples from the experimental room, put them in acontainer that can prevent them from leaking and scattering10) When using human derived samples in the experimental hall (including theexperimental hutch), take necessary measures to avoid scattering of them. In theexperimental hutch, where their leakage may be caused by beam radiation, take necessary
measures to avoid scattering of them.
11) Store solid/liquid waste in a cool dark place after disinfecting/sterilizing or sealing up.

(2) Anonymization
In SPring-8/SACLA, use anonymized human derived samples and data in principle. When using
those that are not anonymized, control them properly to prevent leakage of the personal
information.

(3) Storage
Store human derived samples in dedicatedstorage containers in dedicated storage cabinets,

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Storing human derived samples

August 5, 2016

clearly indicating that the content is such material. If no dedicated cabinets are available, store
them clearly separately from other items. Make sure that you lock the storage cabinets and therooms.
Human derived information must be controlled properly to prevent their theft, loss, leakage andthe like.

(4) Waste
Bring back human derived samples to your laboratory after disinfecting or sterilizing whennecessary. Never leave human derived information at SPring-8/SACLA.

7. Measures to be taken in case of accidents(1) In case of minor abnormalities
When an incident occurs that can be handled by personnel working at the site, follow themeasures shown below:- Judge the situation and follow the measures mentioned in (3) to prevent the effects fromspreading. But take such action only when your safety is ensured.
- Do not handle the incident alone. Always call someone for help.
- Contact the Beamline Toban (PHS: 3899)(2) In case of emergency
When discovering leakage and escape of organisms and the like from experimental facilities
and/or pollution of the human body and experimental facilities by them, or when noticing asituation potentially leading to such incidents, follow the measures shown below immediately.
- When the damage may spread, evacuate first.
- Contact the Beamline Toban (PHS: 3899)- If you need to call an ambulance or a fire engine, inform the Guard House (emergency call: 119
or 0791-58-0828) and they will guide the emergency vehicles. The Guard House will alert thepersonnel and the authorities concerned inside and outside the campus.

(3) When biomaterials are leaked or genetically modified organisms scatter.
When spilling biomaterial, wipe it off to remove. When the material is a genetically modified
organism, inactivate the items and the area contaminated, and for infectious biomaterial, disinfectand sterilize those contaminated.
When experimental animals including genetically modified animals escape within the room,
capture them immediately while taking measures to prevent them from going out of the room.

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August 5, 2016

When experimental plants including genetically modified plants are spread (seed are scattered),recover them promptly.

(4) First aid for injuries that may cause infection (bites/scratches from experimental animals,
contact with pathogenic microorganisms, etc.)
Wash the affected area for 15 minutes or more while squeezing blood from the wound. You can
see a nurse at the Health Office (ext: 3299, 999) near Large Door D3 of the Storage Ring Facility
between 9:00 and 17:30 on week days.
Immediately contact the Health Office (ext. 3299, 999) when an anaphylactic symptom (serious
allergic symptom: hives, breathlessness, chest discomfort, etc.) occurs within half an hour after an
injury. During times when a nurse is absent or outside work hours, contact the Guard House(emergency calls: 119, 0791-58-0828) to request an ambulance. Also inform the Guard House of
the name of the room/building where the injured person is located and describe the injury, which
will be reported to the ambulance on arrival.
Monitor your physical condition carefully for several weeks after an accident to check if suchconditions as high fever, rash, lymph node enlargement and overall malaise occur.
When possible, preserve blood serum of the animal that has caused an infection and when therecords related to infection (quarantine results) are available, check them.

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Manual for biological experiments (second edition)

Emergency calls:
Guard House (ext. 119, 0791-58-0828)
Inquiries:
Safety Office, Japan Synchrotron Radiation Research Institute (JASRI)(Tel: 0791-58-0874, e-mail: safety@spring8.or.jp)
Written by:
JASRI Safety Office
Harima Safety Center, RIKEN