Adipose Tissue

White adipose tissue (WAT)- is largely composed of unilocular lipid-

filled adipocytes that specialize in lipid storage.

Brown adipose tissue (BAT- is largely composed of multilocular

adipocytes that specialize in lipid burning.

-Fat cells appear as “signet rings” on H&E stain because large

lipid droplet displace the nucleus and remainder of the cytoplasm to
the edge of the cell.
-With standard methods of fixation.
-lipids are largely lost from tissues during processing.
-The clear areas in the section are truly empty, because the
triglycerides in the fat droplet are soluble in the organic solvents
used during processing of the section, and are therefore removed from
the section.
-Lipid bodies, also named lipid droplets or adiposomes.
-Cytoplasmic lipid droplets are neutral lipids.
-lipid bodies can be destroyed by drying or fixation and staining
with alcohol based reagents.
-Lipids are difficult to demonstrate, it dissolve in the solvents
used for paraffin processing and, to a lesser degree, in celloidin
processing.
-Frozen sections may be required in order to stain for lipids.
-Triglycerides are always completely removed in properly cleared and
paraffin infiltrated blocks.
-Lipofuscins and myelin may not be completely removed by solvents
used during paraffin processing and significant amounts may still be
present in the tissue sections.
-Lipids are best demonstrated on cryostat sections of fresh unfixed
tissue since there is no really good fixative available.
-Formalin only preserves those lipids that are already more or less
firmly bound to proteins (such as lipofuscins and granules of
leukocytes).
-Lipochrome (lipofuscin) pigments are the breakdown products within
cells from oxidation of lipids and lipoproteins.
-Wear-and-tear pigments found most commonly in heart, liver, CNS, and
adrenal cortex.
-Lipofuscins (lipochrome pigments) are PAS positive and variably acid
fast.
-stain with Ziehl-Neelsen. In addition lipofuscin is Sudan black B
and Sudan Red positive
-Lipochrome can be demonstrated by Schmorl's method which also stains
for melanin.
-Lipochrome may also exhibit a strong orange auto fluorescence in
formalin-fixed, unstained paraffin sections,
-Staining in paraffin sections by exposing the sections to an
emulsion of linoleic acid and lecithin in 70% ethylene glycol at 56oC
for 3 days. These tissues are then treated with 2% chromic acid at
4°C for 24 h followed by 24 h in 5% sodium bicarbonate, with
appropriate rinsing between solutions. Paraffin sections of these
tissues then stained with a lipid-soluble dye such as Oil Red O. The
demonstration of fat embolism with good quality tissue detail is made
practical by the method, which is convenient and inexpensive.
Phospholipids and neutral fats will be lost during routine
dehydration and embedding unless they are treated with potassium
dichromate or osmic acid, which are the only agents that truly fix
lipids. Oxidation of phospholipids by chromate fixation renders them
non-extractable by alcohol, toluene, xylene or paraffin. However,
they both greatly alter the chemical reactivity of the lipids, which
can adversely affect staining. Formol-calcium is the fixative of
choice for lipid histochemistry, and is prepared by adding 2% calcium
acetate to 10% formalin. To preserve the lipids, polyethylene glycols
(carbowaxes) are sometimes used in lieu of the usual dehydration and
embedding process. However, this technique is not widely utilized,
and in most centers, neutral fats are still best demonstrated in
frozen sections of fixed or unfixed tissue. Formol-calcium fixed
tissue blocks are also suitable for cryostat sections, and should be
mounted on chrome-gelatin coated slides, because fixation causes the
endogenous tissue proteins to lose their adhesive properties. In
general, histochemical techniques are the common methods of choice
for demonstrating lipids in tissue sections. These are usually
complemented by other biochemical techniques and chromatography for
specific identification and possible quantification of the lipid
demonstrated by microscopy.