Preservation of fluid specimens prior

to processing
• It is important to preserve the cellular
morphology until the sample can be
processed to get accurate cytologic
interpretation.
• Prefixation: refers to the collection of
specimen in a medium that will preserve
morphology of the cell until the time of slide
preparation
Fresh material
• Specimen may be submitted to laboratories
without preservative if immediate processing
available.
• The length of the time between collection &
preparation of the sample before cellular
damage occurs depends on pH, protein
content, enzymatic activity,& presence or
absence of bacteria
Guide lines for preservation Specimen
• Specimen with high mucus content
• Specimen wit high protein content
• Specimen with low mucus & protein
• Specimen with low pH
 1.Specimen with high mucus content
• Eg: Sputum, bronchial aspirates, or mucocele
fluid
• It can be preserved for 12-24 hrs if refrigerated
• Refrigeration slows down the bacterial growth
which causes the cellular damage & breakdown
of mucus.
• Mucus apparently coats the cells protecting them
against rapid degeneration.
• The cells in specimen without thick mucus or
specimen diluted with saliva are not well
protected & may deteriorate rapidly
2. Specimen with high protein content
• Eg: pleural, peritoneal or pericardial fluids.
• It can preserved for 24 to 48 hrs with
refrigeration
• The protein rich medium act as a tissue
culture medium for the cells & preserve
cellular morphology
3. Specimen with low mucus or protein
content
• Eg : Urine & CSF endure only 1-2 hrs even if
refrigerated.
• The fluid medium in which these cells are
suspended contains enzymatic agents capable
of causing cell destruction.
• Refrigeration may inhibit bacterial growth but
does not protect cells.
 4. Specimen with low pH
• Eg: gastric juice
• It must be collected on ice & be prepared with
in minutes of collection to prevent cellular
destruction by HCl.
 Prefixation of materials
• It may preserve some specimen for days
without deterioration of cells.
• disadvntage:ppt of protein, hardening of cells
in spherical shapes& condensation of
chromatin
 Solutions used as prefixative
• Ethyl Alcohol(ethanol) 50% solution
• Saccomanno's fixative
• Mucolexx
Contin.
• Ethyl alcohol: 50% ethyl alcohol is used
• Ethanol should be added equal volume to the
fluid
• Ethanol in a concentration higher than 50%
should not be used in collecting fluid rich
protein, because the sediment becomes
hardened & very difficult to spread on glass
slides ,particularly if the delay in processing is
greater than one hour.
 Saccomanno’s fixative
• 50% alcohol which contains approximately 2%
carbowax
• Stock solution of carbowax
• 50% ethyl alcohol: 500ml
• Melted carbowax:500ml
• This mixture will not solidify & can be stored
in a bottle.
Contin
• Saccomannos fixative-1L
• 95% ethyl alcohol-526ml
• Stock solution-40ml
• D.W-434ml
• Carbowax infiltrates & occupies
submicroscopic space, preventing cell collapse
& thus protect the cell during air drying.Cell
adhere well to glass slide as a consequence of
air drying.
Mucolexx
• Is a commercial, mucoliquefying preservative
designed for use in the collection of mucoid &
fluid specimen.
• Ingredients are polyethylene glycol, methanol,
buffering agents & aromatics.
• Equal volume of mucolexx & specimen
 Direction for use of fixatives
Preservative Specimen Instruction
Sputum: Have the patient
expectorate directly into
cups containing 50 ml of
50% ethanol preservative & shake well
Saccomanno All body site except gastric
Mucolexx For other specimens add
equal volume of
preservative & mix well

70% ethanol Sputum, bronchial same as above

aspirates& washings

95% ethanol Gastric, bronchi & other Add equal volume of

saline washings preservative & mix well