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Methods To Preserve
Specimens
OUTLINE
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Introduction.
Steps of specimen preservation.
Types of specimens.
Preservatives and their usage.
Vertebrate.
preservation(fishes,herptiles,mammals
and birds
Invertebrate
preservation(arthropods,mollusk,anneli
ds etc)
Precautionary Measures
INTRODUCTION
(a).What Is Meant By Specimen
Preservation?
 Specimen preservation means “longterm preservation of organisms either
plant or animal in the best possible
condition. So that it can be accessed
in future as reference collection for
scientific purposes”
 Many chemical methods are used to
preserve both vertebrate and
invertebrate specimens for
maintainance purposes.
(b).Why specimens are preserved?
Taxonomic reasons.
 For detailed examination.
 For morphological study of particular
animal as each and every animal can’t
be in researcher’s vicinty.
 For zoological museum collection.

Steps For Specimen
Preservation


Killing and relaxing of animal.



Fixation(stops cellular respiration,
kills bacteria within the organism,a
good penetrating ability).



Storage in bottles,jar vials,trays.
Types Of Specimens
(Nagorsen and Peterson, 1980)
1) Entire fluid-preserved animals
Purpose:(for studying anatomy and histology; fluid
preservation may change the fur colour)
2) Study skins with skulls / partial skeletons(some bones in
skin)
Purpose: for studying colour, hair quality and moulting
patterns.
.
3) Mounted skins with partial or entire skeleton (some bones
may remain in the skin, dependant on the method of
preservation) or freeze-dried specimens.

4) Entire skeletons, for instance for studying anatomy,
geographic variation or for age determination.
Preservatives and their usage
 1.


Formalin(Fixative mostly)

Usage:

It is used for vertebrates only.
 It is avoided for long-term storage
since it is acidic and difficult to handle.
 Mostly formalin is used where colour is
important since alcohol dissolves most
colours almost immediately.
 It penetrates more rapidly and internal
organs remain in better condition.

Procedure


Dilution

conc. Formalin(100%) = water saturated with 40%
formaldehyde.

10% formalin = 4% formaldehyde(Used for preservation)

2% formalin with seawater for small specimen.

Mix one part concentrated formalin to nine parts water.

Fill about two-thirds the bottle’s volume with 10%
formalin.

As formalin is acidic, it should be buffered by adding a
pinch or two of sodium bicarbonate.

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

Warning:
Inhalation of formalin fumes is harmful & causes
extreme discomfort to nose and eyes.
Contact with fluid causes severe irritation to the
skin
Contact with sore or raw spots results in extreme
pain.
It is carcinogen.
Hand should be rinsed after usage.
Storage:
It should be kept in safe, water-tight, spill-proof
bottles, e.g. pep-bottles, It should always be
clearly labelled.
2. Industrial Alcohol(for both fixing and
storage)
Usage: Alcohol is usually not used for
killing and fixing vertebrates.But offcourse
used for long-term storage
 Colour of specimen is lost immediately. A
teaspoonful of glycerine in a quart of
alcohol helps to preserve natural
colours and to keep integuments
flexible.
 Alcohol usually comes in the 95%
concentrated form. For long-term
preservation, 70-75% strength is used.

Warning: Alcohol is usually safe to
handle, It can cause irritation to the
skin in cases of prolonged contact.
 Always rinse hands with water after
working with alcohol. Industrial alcohol is
toxic and should never be drunk.
 Alcohol is highly flammable. Never work
with this fluid in the vicinity of open
flames.
 It is rapidly evaporation, and
receptacles holding it should be securely
covered at all times, and not be opened
unnecessarily.

3.Isopropyl alcohol









It is cheap and easy to obtain.
There are different strengths available (70%
and 90%), so if you use isopropyl you want to
dilute it to a 40% alcohol solution.
Isopropyl alcohol can be hard on the
specimens and tends to make them brittle
over time.
Buffering: It can be buffered with
a few drops of glycerin
a pinch of calcium carbonate tablets (crush
the tablets to a powder and add).
4.Ethyl alcohol(for
invertebrates)
The better solution for long term
storage of invertebrate specimens is in
an 80% solution of ethyl alcohol.
 Ethyl alcohol can be found in the
painting supplies.It is labelled as
“denatured alcohol”
 It should also be buffered with
glycerin/antacid tablets.

VERTEBRATES
PRESERVATION


1. Fishes

After capture Fishes are placed in 10% formalin for
quick killing(painful).It is not needed to relax fish
 Fishes dies with its finnature well spread-out, and the
body straight and well-stretched.
 Examination and counting of fin rays and scales quite
easy on such well-preserved material.
 For 30cm fish


i. Formalin - 1 week (fix soft tissue)
ii. Water - 1 day (leach out the formalin)
iii. Alcohol - long term storage
 The length of time for each step may have to be
increased with increasing size of specimens.

2.Herptiles(Reptiles And
Amphibians)
Herptiles are individually kept in plastic & then
killed by freezing or chloroform.
 Shallow trays are used for herptile specimens.
Snakes should be coiled up, and frogs are to be
placed on their bellies with their limbs set at right
angles to their body.
Depending on the size of the specimen, the time
necessary for complete fixation can be different
from 2 days for small salamanders
month for something like alligator snapping
turtle.then added to water for day or two and then in
75% alcohol for long-term storage.

Birds and Mammals:
Usually birds and mammmals are
skinned.
 Skull or Skeletal mounts:
 Flesh can be removed by several means
such as
 Boiling or using dermested beetles.
 Once bones are defleshed they can be
placed in a bleach or hydrogen peroxide
solution to whiten.
 allow to dry; place in bag or box with
complete label tied to skull if possible.

INVERTEBRATE SPECIMEN
PRESERVATION
The easiest way to preserve these animals is
to use alcohol.
 One should be aware of which kind of
alcohol they are are using as each animal
requires a different concentration for
preservation.
 Most invertebrates, however, will be kept in
bottles, and sets of tubes or jars for
preservation
Coelenterates,
Platyhelminthes,Echinoderms










Coelenterates are difficult to preserve.
Preserved in 70% alcohol or 5% formalin.
Hydra can be quickly fixed in Bouin's—warm, not hot.
Platyhelminthes (Flatworms)
Bouin fluid for fixation.
Paraffin , the best long-term preserving and storage medium
of all flatworms.
Echinoderms:
Echinoderms are narcotised by the addition of magnesium
sulphate or menthol to the sea water in which they live.
When completely insensitive to stimuli such as pricking they
should be transferred to 70% alcohol for preservation and
storage.
Arthropods









Easy to process as killed immediately and stored
in alcohol.
Crabs and prawns may also be killed in formalin,
but this renders their joints hard and brittle
The larger arthropods (especially those with
hard exoskeletons) sometimes need to be
injected with 10% formalin to prevent from
rotting.
Industrial alcohol is used for most arthropods.
Insects, crustaceans and arachnids can be
simply dropped into alcohol for immediately
preservation.
It is usually not needed to relax arthropods for
liquid preservation.
Liquid hand sanitizer(for insects)







Hand sanitizer is a gelled alcohol.
Specimens will appear to float in air
inside the vials and do not sink or move
despite any amount of handling.
It is best to kill the animals in an alcohol
solution then transfer them to the hand
sanitizer for preservation.
The gel will break down over time and
become liquid, so you may need to
occasionally replace the gel solution.
Pouring Hand Sanitizer And
Moving Specimen Into Vial
Molluscs





Relaxed mollusk is used for good preservation.
. Newly killed molluscs could then be fixed in
buffered 10% formalin for two or three days,
then transferred to 75% alcohol after soaking for a
few hours in water.
Molluscs can also be relaxed in a solution of
magnesium chloride, but this does not work
very well with land molluscs.
Precautionary Measures






Do not crowd living animals in small
containers - this will result in damage to
their appendages.
Features important in the taxonomic
study of fish, for example, are easily
damaged with contact even after
preservation.
Live crabs before preservation should
be kept individually as some species will
damage each other and other
animals.that will distort morphological
features
Conclusion


. 95% of the museums of the world
use ethanol (drinking or grain
alcohol) for long term preservation.



4.9+% use isopropyl (rubbing
alcohol)



0.001 percent use methanol, or
wood alcohol
Methods to preserve specimens

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Methods to preserve specimens

  • 2. OUTLINE        Introduction. Steps of specimen preservation. Types of specimens. Preservatives and their usage. Vertebrate. preservation(fishes,herptiles,mammals and birds Invertebrate preservation(arthropods,mollusk,anneli ds etc) Precautionary Measures
  • 3. INTRODUCTION (a).What Is Meant By Specimen Preservation?  Specimen preservation means “longterm preservation of organisms either plant or animal in the best possible condition. So that it can be accessed in future as reference collection for scientific purposes”  Many chemical methods are used to preserve both vertebrate and invertebrate specimens for maintainance purposes.
  • 4. (b).Why specimens are preserved? Taxonomic reasons.  For detailed examination.  For morphological study of particular animal as each and every animal can’t be in researcher’s vicinty.  For zoological museum collection. 
  • 5. Steps For Specimen Preservation  Killing and relaxing of animal.  Fixation(stops cellular respiration, kills bacteria within the organism,a good penetrating ability).  Storage in bottles,jar vials,trays.
  • 6. Types Of Specimens (Nagorsen and Peterson, 1980) 1) Entire fluid-preserved animals Purpose:(for studying anatomy and histology; fluid preservation may change the fur colour) 2) Study skins with skulls / partial skeletons(some bones in skin) Purpose: for studying colour, hair quality and moulting patterns. . 3) Mounted skins with partial or entire skeleton (some bones may remain in the skin, dependant on the method of preservation) or freeze-dried specimens. 4) Entire skeletons, for instance for studying anatomy, geographic variation or for age determination.
  • 7. Preservatives and their usage  1.  Formalin(Fixative mostly) Usage: It is used for vertebrates only.  It is avoided for long-term storage since it is acidic and difficult to handle.  Mostly formalin is used where colour is important since alcohol dissolves most colours almost immediately.  It penetrates more rapidly and internal organs remain in better condition. 
  • 8. Procedure  Dilution conc. Formalin(100%) = water saturated with 40% formaldehyde.  10% formalin = 4% formaldehyde(Used for preservation)  2% formalin with seawater for small specimen.  Mix one part concentrated formalin to nine parts water.  Fill about two-thirds the bottle’s volume with 10% formalin.  As formalin is acidic, it should be buffered by adding a pinch or two of sodium bicarbonate. 
  • 9.         Warning: Inhalation of formalin fumes is harmful & causes extreme discomfort to nose and eyes. Contact with fluid causes severe irritation to the skin Contact with sore or raw spots results in extreme pain. It is carcinogen. Hand should be rinsed after usage. Storage: It should be kept in safe, water-tight, spill-proof bottles, e.g. pep-bottles, It should always be clearly labelled.
  • 10. 2. Industrial Alcohol(for both fixing and storage) Usage: Alcohol is usually not used for killing and fixing vertebrates.But offcourse used for long-term storage  Colour of specimen is lost immediately. A teaspoonful of glycerine in a quart of alcohol helps to preserve natural colours and to keep integuments flexible.  Alcohol usually comes in the 95% concentrated form. For long-term preservation, 70-75% strength is used. 
  • 11. Warning: Alcohol is usually safe to handle, It can cause irritation to the skin in cases of prolonged contact.  Always rinse hands with water after working with alcohol. Industrial alcohol is toxic and should never be drunk.  Alcohol is highly flammable. Never work with this fluid in the vicinity of open flames.  It is rapidly evaporation, and receptacles holding it should be securely covered at all times, and not be opened unnecessarily. 
  • 12. 3.Isopropyl alcohol       It is cheap and easy to obtain. There are different strengths available (70% and 90%), so if you use isopropyl you want to dilute it to a 40% alcohol solution. Isopropyl alcohol can be hard on the specimens and tends to make them brittle over time. Buffering: It can be buffered with a few drops of glycerin a pinch of calcium carbonate tablets (crush the tablets to a powder and add).
  • 13. 4.Ethyl alcohol(for invertebrates) The better solution for long term storage of invertebrate specimens is in an 80% solution of ethyl alcohol.  Ethyl alcohol can be found in the painting supplies.It is labelled as “denatured alcohol”  It should also be buffered with glycerin/antacid tablets. 
  • 14. VERTEBRATES PRESERVATION  1. Fishes After capture Fishes are placed in 10% formalin for quick killing(painful).It is not needed to relax fish  Fishes dies with its finnature well spread-out, and the body straight and well-stretched.  Examination and counting of fin rays and scales quite easy on such well-preserved material.  For 30cm fish  i. Formalin - 1 week (fix soft tissue) ii. Water - 1 day (leach out the formalin) iii. Alcohol - long term storage  The length of time for each step may have to be increased with increasing size of specimens. 
  • 15. 2.Herptiles(Reptiles And Amphibians) Herptiles are individually kept in plastic & then killed by freezing or chloroform.  Shallow trays are used for herptile specimens. Snakes should be coiled up, and frogs are to be placed on their bellies with their limbs set at right angles to their body. Depending on the size of the specimen, the time necessary for complete fixation can be different from 2 days for small salamanders month for something like alligator snapping turtle.then added to water for day or two and then in 75% alcohol for long-term storage. 
  • 16. Birds and Mammals: Usually birds and mammmals are skinned.  Skull or Skeletal mounts:  Flesh can be removed by several means such as  Boiling or using dermested beetles.  Once bones are defleshed they can be placed in a bleach or hydrogen peroxide solution to whiten.  allow to dry; place in bag or box with complete label tied to skull if possible. 
  • 17. INVERTEBRATE SPECIMEN PRESERVATION The easiest way to preserve these animals is to use alcohol.  One should be aware of which kind of alcohol they are are using as each animal requires a different concentration for preservation.  Most invertebrates, however, will be kept in bottles, and sets of tubes or jars for preservation
  • 18. Coelenterates, Platyhelminthes,Echinoderms         Coelenterates are difficult to preserve. Preserved in 70% alcohol or 5% formalin. Hydra can be quickly fixed in Bouin's—warm, not hot. Platyhelminthes (Flatworms) Bouin fluid for fixation. Paraffin , the best long-term preserving and storage medium of all flatworms. Echinoderms: Echinoderms are narcotised by the addition of magnesium sulphate or menthol to the sea water in which they live. When completely insensitive to stimuli such as pricking they should be transferred to 70% alcohol for preservation and storage.
  • 19. Arthropods      Easy to process as killed immediately and stored in alcohol. Crabs and prawns may also be killed in formalin, but this renders their joints hard and brittle The larger arthropods (especially those with hard exoskeletons) sometimes need to be injected with 10% formalin to prevent from rotting. Industrial alcohol is used for most arthropods. Insects, crustaceans and arachnids can be simply dropped into alcohol for immediately preservation. It is usually not needed to relax arthropods for liquid preservation.
  • 20. Liquid hand sanitizer(for insects)     Hand sanitizer is a gelled alcohol. Specimens will appear to float in air inside the vials and do not sink or move despite any amount of handling. It is best to kill the animals in an alcohol solution then transfer them to the hand sanitizer for preservation. The gel will break down over time and become liquid, so you may need to occasionally replace the gel solution.
  • 21. Pouring Hand Sanitizer And Moving Specimen Into Vial
  • 22. Molluscs     Relaxed mollusk is used for good preservation. . Newly killed molluscs could then be fixed in buffered 10% formalin for two or three days, then transferred to 75% alcohol after soaking for a few hours in water. Molluscs can also be relaxed in a solution of magnesium chloride, but this does not work very well with land molluscs.
  • 23. Precautionary Measures    Do not crowd living animals in small containers - this will result in damage to their appendages. Features important in the taxonomic study of fish, for example, are easily damaged with contact even after preservation. Live crabs before preservation should be kept individually as some species will damage each other and other animals.that will distort morphological features
  • 24. Conclusion  . 95% of the museums of the world use ethanol (drinking or grain alcohol) for long term preservation.  4.9+% use isopropyl (rubbing alcohol)  0.001 percent use methanol, or wood alcohol