SEMINAR-

TISSUE MICROARRAY

Dr. Nithin
Tissue microarray (TMA)
 The term array generally means:
 When items (such as objects, numbers, etc.) are
arranged in rows and/or columns.
 A tissue microarray contains many small
representative tissue samples from hundreds of
different cases assembled on a single histologic
slide.
 Which allows high throughput analysis of multiple
specimens at the same time.
Small representative tissue samples from hundreds of different cases assembled on a single
histologic slide.
INTRODUCTION
 TMA helps to perform immunohistochemistry (IHC),
fluorescent in situ hybridization (FISH) or RNA
analysis for hundreds of cases in a single slide.
 Therefore the technique is faster and cheaper than
the conventional method.
 Many studies on human cancer have identified
multiple novel markers, primarily at the gene level,
which has a prognostic, diagnostic or therapeutic
significance.
INTRODUCTION
 The validation of these markers using the standard
histopathological techniques is time consuming, and
labor intensive and costly, particularly when
multiple markers are tested on numerous specimens
 Tissue microarray is a recent innovation in the field
of pathology that is expected to overcome these
significant problems.
Tissue Microarray Technique
 At first, H&E -stained section from the donor block is
studied, and the representative area of the donor
block is marked.
 With the help of tissue microarray instrument, core
tissue biopsy (0.6–2 mm diameter) is taken from the
donor paraffin block and is placed in an empty
paraffin block (the recipient block) in precise
manner
 0.6 mm core biopsy = 2-3 HPF of tissue = 600
cases/ slide.
Tissue Microarray Technique
 The recipient block can accommodate few hundreds to
thousand specimens ( based on the diameter of core
biopsy).
 The co-ordinates of the core biopsies in the recipient
block are recorded typically in a spreadsheet
(preferably in Microsoft Excel file).
 Recipient block is heated at 40 degree for 15 min until
the surface becomes flat.
 Now with the help of a microtome, 3-5 μ sections are
cut from the TMA block to produce TMA slide
 The section now can be used for IHC, FISH or other
molecular studies.
Tissue Microarray Technique
 Number of Cores in TMA In a 2.5 × 4.5 cm block, at
least 1000 cores can be adjusted. However it is
preferable not to take more than 500 cores in a single
block.
 Automated TMA Presently automated tissue microarray
machine is available that bypasses the tedious manual
punching procedure.
 It helps to design the array design or layout, and the
machine automatically creates holes in the paraffin wax
and places the donor tissue in the hole of the recipient
paraffin wax.
Advantages of TMA
 Amplification of the resource:
 Ordinarily from a standard 5 mm thick section of tissue,
we can get maximum 100 sections for study.
 Whereas the microarray technique could produce
material for 500 000 assays (assuming 250 biopsies
per section times 2000 2.5-μm sections per 5 mm array
block) represented as 0.6 mm disks of tissue.
 Thus this technique essentially amplifies (up to 10 000
fold) the limited tissue resource
Advantages of TMA
 Simultaneous analysis of very large numbers of
specimens
 Uniformity in staining conditions:
 In conventional staining, slide-to-slide variation may
occur due to several variables such as antigen retrieval,
concentration of different reagents, incubation period,
washing time, etc.
 However in case of TMA, each tissue section consists of
100–1000 core biopsies from the different patients,
and the single section is stained that avoids all the
slide-to-slide variability.
Advantages of TMA
 Faster, cheaper and reduction of manpower:
 In TMA a single slide requires less reagents, labor and
time. Therefore, TMA saves cost, time and total work
force.
 Original block can be preserved:
 Only a few core biopsies from the original block are
sufficient to make TMA.
 The original block can be preserved and can be used
for further sectioning.
Advantages of TMA
 Effective in quality assurance program:
 Due to significant amplification of the laboratory
material, TMA can be used for external and internal
quality control program.
 The TMA section can also be used for standardization
of reagents for positive control.
Limitations of TMA
 Tissue heterogeneity:
 This is one of the main concerns of TMA.
 The tumor may vary from area to areas such as Hodgkin
lymphoma which may have different morphologies in
different areas.
 Therefore small 2 mm tissue may not represent the whole
tumor and finding may vary.
 However, several studies have shown that TMA and whole
tissue data are almost similar.
 Less cost-effective in small series of cases:
 TMA is not very effective if it is done once in a while in a
small series of cases.
Limitations of TMA
 Prone to loss the tissue:
 The core biopsy tissues may be lost due to its tiny size.
 Tissue rich in keratin, bone or cartilages are likely to be
lost.
 Disorientation of the core biopsy tissue:
 Due to large number of core biopsies, there is a chance
of disorientation when TMA is done manually.
 Rows of empty core tissues may help in the immediate
orientation of the tissue.
IMPLICATION IN RESEARCH

 Tissue microarray facilitates rapid translation of

molecular discoveries to clinical applications.
 Newly identified genes require clinical validation on
histopathological specimens for any meaningful
outcome.
 Such validation is best achieved by tissue
microarray
 The technique has been applied to tumor research
(gliomas, breast tumors, lung cancer among others).
IMPLICATION IN RESEARCH
 three categories of tissue microarray can be
defined:
 multitumor arrays ( example expression of PDL1 in
gastric, bladder, colonic and breast carcinoma)
 tumor progression arrays ( example expression in
AMACR in Normal prostate, BPH, PIN and different
stages of prostatic carcinoma)
 prognostic arrays
Conclusion
 Tissue microarray is a practical and effective tool
for high-throughput molecular analysis of tissues
that is helping identify new diagnostic and
prognostic markers and targets in human cancers.
 It has varying degrees of research use and offers a
range of potential applications in basic research,
prognostic oncology and drug discovery.
 Tissue microarray technique will lead to a significant
acceleration in the transition of basic research
findings into clinical applications.
THANK YOU