RUNNING HEAD:

Name of the Student

Name of the university

Table of Contents
Introduction:.........................................................................3
Methodes and Material:.............................................................4
Discussion:...........................................................................4
Investigation & Diagnosis:..........................................................6
Recommendation:...................................................................7
Conclusion:........................................................................... 7

Introduction:
Just below the rib cage are the two bean-shaped organs called kidneys. Kidneys in
human body performs one of the most important function- the blood filtration and
waste product excretion. Glomerulus synonymous with filtration are tiny clusters of
blood vessels present inside the kidney in which blood enters via branching out
arteries. One kidney has approximately 1 million glomeruli in it. With Glomerulus
(one glomeri) is attached a small fluid collecting tube called tubule. After the blood
filtration in glomerulus the extra wastes and fluid passes through tubule and becomes
urine. And finally, the kidneys through a large called ureter extreme the urine into the
bladder.
A branch of pathology, Histopathology is a specialized study of the disease of the
tissues by definition. Before the microscopic diagnosis of the tissues they must pass
through a number of steps like fixation, embedding and staining. In order achieve a
more correct diagnosis there is a need of a lot more accurate preparation such as
enough fixation time, sufficiently thick or thin section, good quality staining and well
embedding. Consequently, all the architecture will be well differentiated and clear.
Moreover, H&E stain, special stain, in situ hybridization and immunohistochemistry
are some of the different kinds of methods in Histopathology for diagnosis.
Furthermore, stained tissues investigations through microscope is one of the best
methods to get rapid and correct diagnosis. Besides using H&E, which is the routine
stain, through this report, in the given tissue section we will try to identify and
diagnose the abnormalities by using different kind of special stain. The microscopic
examination result will help in finding the exact reason of patient symptoms.

Methodes and Material:

Along with clinical history of the case study, H&E section was given. After
microscopic examination of H&E section some disease has been assumed. Hence to
identify the exact type of the disease, five special stains were taken into consideration
to be performed on five unstained slides. First of all by using xylene the five slides
along with five control slides were de-waxed and then hydrated by descending
alcohols and further running tap water. Secondly, using different special stain as
labeled each slide was stained. In the third step, by ascending alcohols all the sections
were hydrated except that of gram stain slides, which were air-dried, then all the slides
including gram stain slides were cleared with xylene. Further mounted by the use of
DPX and then labeled. Finally, all the slides were microscopically examined and the
results were recorded.
Discussion:
Kidney in humans is one of those organs, which have too many diseases associated to
it such as renal cell carcinoma, pyelonephritis and glumurlnephritis. Therefore it
requires a lot of experience to diagnose the correct disease. In our case, some of the
glomeruli are showing as hypercellular and hyalinized in the H&E section, but it can
be false appearance also as it is difficult to differentiate between mesangial cells and
endothelial. In addition to that, there is presence of inflammatory cells in the H&E
section. This could point towards inflammation of the kidney. Further, the patient is a
young woman and looking at other clinical features like hematuria with pain and since
women is at higher risk of kidney infection because of a shorter urethra compared to
men, so in my initial diagnosis I suspect the patient has pyelonephritis. In order to be
sure in diagnosing the disease and eliminate some of them it is suggested five special

stains to be used. Those special stains are Masson Trichrome, Periodic acid Schiff,
Gram Stain, and Ziehl-Neelsen method for acid fast bacilli and Grocott Methinoamne
Silver. First I have chosen Gram Stain as it can distinguish between negative and
positive gram bacteria. Some bacteria like E. coli or klebsiella through lower urinary
tract system can reach kidney and are most causative bacteria for pyelonephritis, so in
order to identify gram stain is needed and distinguish this type of bacteria if present.
Secondly, in order to eliminate the possibility of renal tuberculosis Z-N stain were
suggested to identify the presence of mycobacterium species that can cause the
disease. It is possible that bovine tubercle bacillus is responsible for the disease but
M. tuberculosis is the most common causative agent. Therefore to identify if
tuberculosis is present or not Z-N stain is important to be performed. Thirdly, the
GMS stain which demonstrates the presence of fungi. Cryptococcus, species candida
and aspergillus are the fungal infection which have been reported to cause renal
parenchymal, since the use of immunosuppressive therapy with organ transplantation
has been increased. Fourthly, in renal biopsy to demonstrate and evaluate the amount
of fibrous connective tissue the stain Masson Trichrome were suggested. In addition
to that it will help in distinguishing between interstitial fibrosis and interstitial
oedema. Furthermore it can easily calculate the lobular accentuation of the glomerular
tufts and bowmans capsule. Lastly, the stain PAS stain was suggested to evaluate the
glomerulus as it is the best to do so and moreover it demonstrates bowmans capsule
and glomerulus basement membrane. Furthermore, PAS stain can easily picture the
spread GBM thickening that is seen in many glomerulus infections such as diabetic
glomerulopathy, hypertension and membrane glomerulonephritis. Moreover, PAS
stain positively for sclerosis, hyaline deposits, proximal tubules and mesangial matrix.

Therefore it is important to order PAS stain so as to correctly evaluate and

demonstrate the most of kidney architecture.
Investigation & Diagnosis:
Since the Z-N stain, gram stain and GMS are negative it can be concluded that no
microorganism infection present in the kidney. Consequently as expected, fungal
infection, pyelonephritis and renal tuberculosis are eliminated from the diseases list.
But on the other hand fibrosis in the tissue is shown by masson trichrome which
demonstrates the fact that infection is present in the kidney. It should be noted that if
excessive collagen is present in the tissue, it suggests that the tissue is recovering from
the chronic diseases. So clearly in this case, the kidney is regenerating after injury.
Additionally the diffuse thickening of the glomeruli capillary wall was shown by the
PAS stain. Glomeruli showed normal glomeruli, segmental scleroses are present
though. Caused by the obstruction of blood flow there is presence of tubules with
thick redundant basement membrane (tubular atrophy). Moreover, some of the
bowmens capsule and glomeruli are replaced with a pink in color, homogeneous and
amorphous material called hyaline (hyalinosclerosis) that is resulted from
combination of increased collagen, plasma protein and mesangial matrix. Therefore
depending on the clinical features and the given microscopically diagnosis the patient
seems to have focal segmental glomerulosclerosis (FSGS). In adults, focal segmental
glomerulosclerosis is one of the most possible causes of primary glomerular diseases.
It is not necessarily a distinct disease and represents characteristic pathologic pattern
of glomerular injury. The increased degree of scarring of some glomeruli (focal) in
kidney biopsy is a sign for (FSGS), also only a portion of glomeruli in involved in the
scarring dose. However, some normal glomeruli are also present which can be

classified to collapsing glomerulopathy like primary (idiopathic) and HIV infection

like secondary. In adult this condition is considered to be the predominant cause of
idiopathic neghrotic syndrome and within 10 to 20 years, 40% to 60% of patients
move further to end stage renal disease (ESRD).
Recommendation:
To confirm the early diagnosis further tests should be done. In order to detect early
signs of kidney damage, a urine microalbumin test should be done which will measure
the protein amount present in the body. Additionally, in order to calculate the removal
of creatinine, which is the waste product from the kidney, the glomerular filtration rate
(GFR) is recommended. The above-mentioned two tests are useful in diagnosing the
presence of lubus nephritis and diabetic nephropathy. Furthermore, it is
recommended that some other special stains performed, for instance, to demonstrate
amyloidosis, Congo red to be performed. Lastly, AE1/AE3 is an example of antibody
and it is important to perform immunohistochemistry to diagnose renal cell
carcinoma.

Conclusion:
To discover most of the diseases it is essential to perform microscopically diagnosis.
However, quality in tissue processing in needed for correct diagnosis. Therefore it is
necessary to have quality control in histopathology section. As there are numerous
Kidney diseases, therefore there is a need of several tests in renal biopsy and special
stains to get the correct diagnosis. Further, In this case, different results shown by
special stains aided in eliminating some of the diseases and helped in giving the
correct diagnosis. While special stains for microorganism gave the negative result,
masson trichrome and PAS gave positive result. Therefore, it can be concluded that

the patient suffers from the glomerular disease specifically that of FSGS (focal
segmental glomerulosclerosis). Finally, other diseases like amyloidosis, diabetic
nephropathy and renal cell carcinoma are suspected too, but in order to confirm them
they need to be tested further.