Preparation of Slide Tissue

Prof. Dr. Diyaa Aljaza

Anatomy Department
 Tissue preparation
1) Collection samples.
2) Fixation tissue.
3) Washing tissue.
4) Dehydration.
5) Clearing.
6) Embedding
7) Frozen tissue.
8) Sectioning .
9) Staining technique.
 Collection of tissue
 Take fresh or perfused tissue and cut into 1
cm cube and keep with 10% formalin.

 For example of tissue:

Uterus, lung, intestine, colon, breast, stomach,
skin, thyroid, liver, pancreases…….ect.
 Fixation
 Tissue keeped in container with 10% formalin
for 24 hr at least until one year.

 The purpose of fixation is to preserve the

tissue structure.
 Washing
 Tissue sample wash with tap water four times.

 The purpose of washing is to remove excessive

formalin from the tissue.
 Dehydration
• The tissue sample is soaked in a series of
alcohol concentrations in an ascending model
(30, 50, 70, 80, 90 and 100%) for two hrs for
each step.

• The purpose of this step is to remove all

fixating solution and tissue fluids.
 Clearing
 Sample is soaking in pure xylene solution.

 This step replicates twice 1 hr for each.

 The purpose of this step is to remove the

excessive alcohol.
 Embedding
• This process is performed by using paraffin wax at
60oC for 6 hrs.

• In this step the wax will infiltrate in side tissue

and replace the wax.

• Frozen the wax blocks

• The purpose of the step is to remove xylene and

provide external support.
 Sectioning
 By using rotary microtome, paraffin blocks sectioned
into sections (3-5 µm thick).

 Float the tissue section in warm water bath at 40oC to

flatten.

 Pick up sections onto glass microscope slide coated

with Mayer’s albumin.

 Produce sections thin enough to allow viewing through

a microscope
 Staining
• The purpose is to give color and contrast to tissue
structures.
• Dewaxing using xylene tiwicely for (3 min) and
rehydration by socking into a descending series of
ethanol (100-70%, respectively) for two min each
time.
• Rinse in D.W for 2 min.
 Haematoxylin and Eosin stains
• Tissue slides dyed with Haematoxylin stain for 10
min and then wash with D.W. to stain nuclei blue.
• The second stain is Eosin to stain the cytoplasm.
• Some drops of Disteren Plasticizer Xylene (DPX)
place on the slide and cover with coverslips (22 x
22 mm).
• Left to dry and next day the slid ready to examine
under light microscope 40x magnification powers.
 Reference
• Luna, L.G. (1968).
 Thank you

For your attention