m
 

Õ %
  is a chemical process by which biological

tissues are preserved from decay, either through
 or
%  . Fixation terminates any
ongoing biochemical reactions, and may also increase
the mechanical strength or stability of the treated
tissues
Õ fixative usually acts to disable intrinsic biomolecules Ȃ
particularly proteolytic enzymesȄwhich otherwise
digests or damages the sample.
Õ a fixative typically protects a sample from extrinsic
damage
Õ fixatives often alter the cells or tissues on a molecular
level to increase their mechanical strength or stability.
Ú



Õ %
 
Õ  
 
    %
   
Formaldehyde fixes tissue by cross-linking the
Proteins
Õ ñlutaraldehyde is a larger molecule, and so its rate of
diffusion across membranes is slower than
formaldehyde.
Õ One of the advantages of glutaraldehyde fixation is
that it may offer a more rigid or tightly linked fixed
product.
Õ Its greater length and two aldehyde groups allow it to
'bridge' and link more distant pairs of protein
molecules
Õ It causes rapid and irreversible changes, fixes quickly,
is well suited for electron microscopy, fixes well at 4oC,
and gives best overall cytoplasmic and nuclear detail.
Õ ©
 %
    
Precipitating (or •

) fixatives act by
reducing the solubility of protein molecules and
(often) by disrupting the hydrophobic interactions
that give many proteins their tertiary structure.

 %
  can react with various side
chains of proteins and other biomolecules, allowing
formation of crosslinks that stabilize tissue structure
Õ ©  penetrate tissue well to react with histones
and basic proteins to form crystalline picrates with
amino acids and precipitate

  %%  
 

©
Should be kept in the physiological range
between pH 4-9.
Õ 
 
d hypertonic solution contains a greater concentration
of impermeable solutes than the solution on the other
side of the membrane. When a cellǯs cytoplasm is
bathed in a hypertonic solution the water will be
drawn into the solution and out of the cell by osmosis.
If water molecules continue to diffuse out of the cell, it
will cause the cell to shrink, or crenate. solution having
greater solute concentration than the cytosol.
Õ 
 
d hypotonic solution contains a lesser concentration of
impermeable solutes than the solution on the other side of
the membrane. When a cellǯs cytoplasm is bathed in a
hypotonic solution the water will be drawn out of the
solution and into the cell by osmosis. If water molecules
continue to diffuse into the cell, it will cause the cell to
swell, up to the point that lysis (rupture) may occur.
Õ %
 
dt least 15-20 times greater than tissue volume.
Õ Ú

Increasing the temperature increases speed of
fixation. However, care is required
Õ 0