Collection and storage of tissues

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Collection and storage of tissues
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Regulations governing acquisition of specimens
Collectors should become familiar with local,
state, national, and international laws and
regulations, and they should allow adequate lead
time to obtain the necessary permits.
Regulations concerning the collection and
transport of biological materials are designed to
safeguard the general public health, protect
domestic crops and livestock, and control illicit
traffic in endangered and threatened species.
Scientists intending to import frozen tissues
and other specimens must adhere to all applicable
wildlife regulations for the countries
involved. Scientific collecting permits usually
are necessary for sampling natural populations
these are obtained from state and national fish
and wildlife forestry or conservation offices.
Six months may be required to obtain permits
for collecting this may be extended to a year or
more if international travel or endangered
species are involved.
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Different documents may be required to collect,
to travel in certain areas, to export from the
country of origin and to import into the
researchers country. To verify the animals
good health, some countries require a quarantine
period following importation of certain living
organisms.
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Removing and preserving tissues in the field
The collection and handling of tissues for use in
future molecular studies can be carried out by
individuals with minimal training. Tissues
should be sampled while the organism is alive or
as soon after its death as possible. Even a
tiny amount of tissue is valuable. The kinds
and quantities of tissues preserved depend on the
needs of the individual investigator however we
recommend that collectors maximize diversity and
quantity of tissue types to help develop synoptic
collections. Collectors should be aware of the
importance of keeping instruments, containers,
and reagents clean, and of placing tissue samples
in a cold environment and away from light as
quickly as possible. Tissues should be packaged
in plastic cryotubes, plastic bags or wrapped
tightly in aluminum foil.
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• Packages should have space for tissue expansion
  during freezing but otherwise have as little
  unused space (airpockets) as possible to minimize
  drying of tissues and denaturation of
  macromolcules.
• Careful documentation of samples is critical in
  all phases of work. A sample in a cryotube or
  other package is essentially useless if it has
  lost its label.
• It is important to
• Label samples and specimens so that no
  information is lost in wrapping, transport,
  storage and entering of data into permanent
  records
• Cross-reference the tissue sample with field
  collection data for the original specimens
• Label containers, laboratory notebooks and
  experimental samples during study
• List specimens examined in research publications.
  Ideally this citation will include the museum
  catalogue number for the voucher specimen housed
  in a permanent repository. Although the museum or
  herbarium number may be assigned long after the
  tissue sample was collected, all records
  pertaining to the sample (e.g., field catalogue
  data, notes and photographs) should be
  cross-referenced with the permanent voucher
  number.

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We recommend that individuals collecting tissue
specimens in the field continue to use
traditional collectors catalogues. These
usually are organized so that each specimen
receives a unique number preceded by the
collectors initials. The catalogue entry
should indicate the types(s) of tissues sampled.
The package or tube containing the tissues
should be marked clearly with the collectors
initials and field number. The name of the
specialist who provided identification of a
specimen is and important part of the
documentation. Great care should be taken in
labeling tubes and packages containing tissue
samples. The following items have been
reliable 1) high-quality bond paper and a
drafting pen with permanent, waterproof,
non-smearing ink 2) felt-tip pen with permanent
ink that adheres to plastic tubes or packages
3) a ballpoint pen that leaves a clear
impression in aluminum foil.
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Experiment by exposing your material to liquid
nitrogen or other ultracold conditions followed
by thawing. In addition, we recommend use of a
backup system (e.g., number written in ink and
also etched on the tube with a diamond-tipped
scribe labels both inside and outside the
package). On occasion, it is not possible to
cross-reference a tissue sample with a
permanently preserved voucher specimen, such as
when a blood sample is taken from an individual
in a zoo of from one that will be released after
temporary restraint in the field. Under such
circumstances, photograph the individual to
document its identification and record its tag,
band or other identifying number, if known.
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Procedures unique to animal tissue collection
Animals collected should be handled with care
consistent with appropriate guidelines for their
welfare. We recommend that tissues be collected
and packaged individually when larger animals are
sacrificed. With vertebrates, blood, heart,
liver, kidney, stomach, intestine, a sample of
skeletal muscle and perhaps other organs should
be collected. Red blood cells of al vertebrates
except mammals and some amphibians are excellent
sources of DNA the buffy coat of white blood
cells is a good source of mammalian DNA.
Valuable material for molecular studies such
as blood, hemolymph, snake venom, feather pulp
and muscle biopsies can be obtained without
sacrificing the animal.
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Transport of tissues from field to laboratory or
between laboratories
Tissues are usually transported either in
Styrofoam boxed packed with dry ice or in liquid
nitrogen containers. Small samples are
conveniently carried in personal baggage in
thermos bottles containing refrigeration packs.
Dry ice and liquid nitrogen are classified as
restricted articles by the International Air
Transport Association, and the shipper must be
aware of all pertinent regulations. Dry ice
containers are usually accepted as baggage by the
airline agent if the Shippers certification for
restricted articles is attached to the package.
No more than 200 kg of dry ice may be shipped in
a single package. Liquid nitrogen in
nonpressurized, metal dewar flasks is also
authorized for shipment by air. No more than 50
kg per flask can be shipped on an aircraft
carrying passengers.
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The majority of tissue samples are stored in an
electric ultracold freezer (-70 to -150ºC) or in
liquid nitrogen. Ease of access of samples
within the freezer or nitrogen tank is of special
importance. Samples can be stored in numbered
moisture-proof boxes. Ultracold storage space
is very expensive to purchase and maintain, and
it is important that materials be stored in a
space-efficient manner. Thus, access and
inventory procedures for frozen tissue
collections should be extremely well organized.
Freezers should be opened as rarely as
possible ultracold freezers are sensitive to
even brief periods of temperature warm-up. One
must know exactly where each sample is located
before opening the freezer. Freezers should be
monitored at least once each day and should be
equipped with both local and remote systems that
will sound alarms in the event of electrical or
mechanical failure. Ideally, liquid nitrogen is
better than ultracold freezers for long-term
storage because of its much colder temperature
(-196C).
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Biobank
Biomaterials derived from wildlife in Southern
Africa represent a valuable natural resource
belonging to its people. Biomaterials include
all tissue, fluids and genetic material derived
form wildlife such as skin, feathers, blood and
blood products, sperm, muscle, egg-cells, horn,
embryos and hair which may be used for wildlife
conservation , research and management. Most
conservation actions today involve the direct use
of, or knowledge derived from the analysis of
wildlife biomaterials. Biomaterials and the
associated information are relevant to those
working in the fields of genetics, molecular
biology, disease diagnosis and management,
epidemiology, animal physiology, reproduction,
taxonomy, wildlife and small population
management, legislation and policy, forensics and
law enforcements, nutritional studies and so on.
With the advancements made in biotechnology in
recent years, the options for collecting, banking
and utilizing biomaterials for effective
conservation have increased enormously, as have
the institutions and organizations involved in
this aspect of wildlife management and
utilization.
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A biobank is a system for the management of a
tailored facility that collects, stores and
distributes biological resources. This system
incorporates an adaptive data model and developed
processes for the management of intellectual
property, policy issues, services and
maintenance. The following issues need to be
considered when banking tissues.
Which tissue do you bank? How many do you
bank? Which species do you bank? How do you store
the tissue? Where do you bank the
samples? Disease issues? Database and information
technology? Biobanking as a potential the key to
conserving the worlds biodiversity and will
contribute to new discoveries in conservation,
medicine and agriculture.
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