TISSUE MICROARRAY
By:
Dr. Salonee Rajdev
2nd year resident
WHAT IS TISSUE MICROARRAY
• Tissue microarray is a recent innovation in the field
of Pathology.
• A microarray contains many small representative
tissue sasmples from hundreds of different cases
assembled on a single histologic slide.
• It allows analysis of multiple specimens at the same
time
• This is done by using a needle to biopsy a histologic
tissues and placing the core into an array on a
recipient paraffin block.
HISTORICAL ASPECT
• They were initially designed to compare many
different tissues or cells in one slide and to
provide a cost effective procedure which saves
the staining consumables.
• Origin by : Dr. Hector Battifora, 1986
• Term Tissue Microarray was coined by
Kononen et al in 1988
CHARACTERISTICS AND ADVANTAGES OF
TMA
• 50-500 tissues can be analyzed per block
• High throughput
• Relatively lower cost
• Can be used for a wide variety of staining
procedures including :
IHC ISH FISH H&E
SPECIAL STAIN
CONTROL SAMPLES
TYPES OF TMA
• Based on material of origin :
PARAFFIN TISSUE
FROZEN/CRYOARRAY
ARRAY
Consists of TMAs from frozen Consists of TMAs constructed
tissue . from paraffin embedded
The DNA, RNA and protein in material.
frozen tissues retain their native
characteristics.
TYPES OF TMA
• Based on their application or the question being asked in the study, viz.
1. Cryo-TMA
2. Multitumor TMA : many different types of tissues aligned on the slide.
3. Progression TMA : different stages of tumor/disease progression within a given
organ.
4. Prognosis TMA : Disease samples/ tumor biopsies taken from patients, examined
and analyzed with clinical data to come to a prognostic aspect of the disease. Can
be used with a progression TMA.
5. Cell line TMA : Consist of normal or cancer cell lines that are grown in culture. The
major function of this to assess the presence of proteins that are known to be
present in one or more cell lines.
6. Random tissue array
7. Consecutive case array
8. Outcome based TMA : Most valuable and most difficult to generate as they involve
collation of tissues from patients that have more or less similarly treated and
followed up for a significant period of time.
DESIGN AND CONSTRUCTION OF TMA
• In the tissue microarray technique, a hollow needle is used to
remove tissue cores as small as 0.6 mm in diameter from regions
of interest in paraffin-embedded tissues such as clinical biopsies or
tumor samples. These tissue cores are then inserted in a recipient
paraffin block in a precisely spaced, array pattern. Sections from
this block are cut using a microtome, mounted on a microscope
slide and then analyzed by any method of standard histological
analysis. Each microarray block can be cut into 100 – 500 sections
• STEP 1: TISSUE COLLECTION
At first, retrieve the stored paraffin donor blocks and are sectioned
to produce a standard histopathological slide stained with H&E.
• STEP 2 : MAPPING OF SLIDES
Map the area of interest on a fresh H&E slide on
a scanner view with ink.
Then the block is usually matched to the
corresponding glass slide.
Donor blocks must be atleast 1mm thick to be
suitable for array construction.
STEP 3 : DESIGNING THE GRID
It is important to plan and record in advance
how many samples will be arrayed.
And to create a map or grid sheet which will be
easy to follow and refer to later.
STEP 4 : PREPARATION OF RECIPIENT BLOCKS
Different sized moulds are available for
preparation of recipient blocks.
Size of the punch is important ( 0.6,1.0,1.5,2.0
mm)
STEP 5 : ARRAYING OF DONOR BLOCKS
Manual Arrayer : a manual arrayer relies on the
designed grip map sheet and direct identification
and manual selection of the punching is done. (~30-
70 cores/hour)
Portable quick arrayer : Portable, easy to handle and
economical. Needles are replaced by tips.
Automated arrayer : Includes a specimen tracking
software. Ideal for a lab with high volume of TMA
(~120-180 cores/hour)
TROUBLESHOOTING IN TMAs
• 1. Core doesn’t come out of the punch easily –
suggests that the punch is bent/distorted.
Solution : Change the punch.
• 2. Tissue core was pushed too deep – Remove
the sample with a small punch and put a new
sample in the same position.
• 3. Insufficient spacing of cores – can cause
minor cracks while sectioning
TROUBLESHOOTING IN TMAs
• 4. Thinning of TMA cores in block – can lead of
unevenness of a core in the block and renders
the block unusable.
• 5. Double stacking is also advised to make sure
target tissue is present in more sections of the
TMA.
APPLICATIONS
• Validation of diagnostic biomarkers
The most popular use of TMA is in basic/translational
research in validation of diagnostic markers in annotated
clinical samples. Given the rapid pace by which new genes
are being discovered by high-throughput DNA microarrays,
TMAs hold an immense potential to validate this genomic
and proteomic data across multitumor types in a limited
timeframe.
Diagnostic biomarkers are tissue markers that can serve as
screening tools for certain diseases. or for identification of
specific subclass in a disease.
• Validation of prognostic biomarkers
TMAs are becoming the standard for the validation of prognostic
biomarkers. As tumor banks and clinical sample cohorts are
maturing with regard to clinical follow-up data, it is becoming
possible to perform correlations of biomarker expression with
clinical endpoints like disease-free survival or overall survival.
• TMA and response to therapy
TMAs are of increasing utility in defining the feasibility of
developing particular therapeutic targets. They are now being
constructed from samples of patients receiving specific
treatments providing a unique tool for validating predictive
response to these treatments.
• Non-neoplastic TMAs
Increasing use of TMAs for research in neurdegenerative
diseases, dermatological and cardiac diseases.
• Clinical applications
TMAs are being used for clinical applications in Pathology departments for
testing of new antibodies and probes, or determining optimal staining
conditions using small test TMAs
Application of TMA is emerging in clinical trials where the TMA is
constructed from pathology tissues taken before initiation of therapeutic
trial. The objective is to define the functional status of the tumor prior to
therapy.
• Quality control (QC)
Immunohistochemistry analyses have been justly criticized for their
subjective and semiquantitative means of determining the level of protein
expression. QC in IHC is one of the major problems in diagnostic pathology.
LIMITATIONS
• The most obvious drawback of this technology was initially
believed to be the possibility that small tissue cylinders
might not adequately represent the whole sections,
particularly in case of tumors, because of intratumor
heterogeneity of protein expression. Thus, increasing the
number of cores collected from the sample and/or
increasing the core diameter to 1.0-2.0mm can resolve this
limitation.
• A tumor tissue may comprise of many different histological
areas within itself, such as regions of apoptosis, necrosis or
increased proliferation and it may not be possible to sample
all areas in one tissue. Hence, care should be taken while
taking the cores from the original tissue and focus should
me made on the purpose of the investigation.