Name: modified pasm-masson staining in a renal biopsy
Uploaded: 2019-07-24T11:27:04
Description: Special histochemical staining is an important method for a renal biopsy, which includes PAS staining, periodic acid silver methenamine (PASM) staining, ...

This work was supported by National Natural Science Foundation of China (Grant No. 81872202); by the Natural Science Foundation of Guangdong Province (Grant No. 2015A030313263, 2018A030313778); by Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research (Grant No. 2017B030301018); by the Natural Science Novel Project of Department of Education of Guangdong Province (Grant No. 2013KJCX0093).

The protocol follows the guidelines of our hospital&#39;s human research ethics committee. One hundred and fifty-two cases of renal biopsy specimens were collected from the Department of Pathology at the Shenzhen Third People&#39;s Hospital (The Second Affiliated Hospital of Southern University of Science and Technology) from January 2018 to May 2019.
1. Material preparation
<ol>
	<li>Fix all tissues with 4% neutral buffered formaldehyde (pH 7.0) for 24 h.</li>
	<li>Dehydrate the fixed tissu...

<ol>
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Pathological diagnosis is an important tool for the diagnosis and differential diagnosis of renal diseases, and it is of great significance for the guidance of treatment and predicted value of renal diseases10,11,12. Special staining is an essential method for renal biopsy including PAS staining, PASM staining and Masson staining3,13,14...

Special histochemical staining has been used by pathologists to assist in tissue-based diagnoses for over 100 years1,2. The application of special staining in the diagnosis of renal diseases has been recognized as an important method for distinguishing the types of renal diseases and displaying some special components in renal tissues3.
To date, a variety of special stains have been applied to renal biopsies, which includes periodic acid silver methenamine (PASM) staining and Masson&#39;s trichrome staining. Lesions of the glomerular capillary walls are the main pathological changes, which helps to characterize and differentiate various glomerular diseases. PASM staining is mainly used to observe the degree of abnormalities of the glomerular capillary walls in glomerular basement membrane4,5. Masson staining is utilized to investigate an extremely wide variety of glomerular diseases6,7 and has been found to be effective in identifying the accumulation of the immune complex as well as the proliferation of basement membranes and renal interstitial collagen fibers8. For example, chronic graft damage in both the medulla and the cortex in renal transplant biopsies was measured by PASM and Masson histochemistry, respectively9. However, if PASM and Masson staining are done separately, it is difficult to accurately detect the localization of the basement membrane and immune complex, and to judge whether these two lesions exist at the same site. On the contrary, PASM-Masson dual staining can clearly distinguish the staining and localization of the basement membrane and the immune complex in the same section. Moreover, it can shorten the staining time and improve the work efficiency. Therefore, PASM-Masson staining plays an important role in the diagnosis of glomerular diseases.
In order to identify the location and extent of glomerular immune complex deposition in different types of renal diseases, 152 cases of renal biopsy were collected and PASM-Masson dual staining was performed in the present study.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Absolute ethyl alcohol</td>
			<td>Guanghua company (Guangzhou city, China)</td>
			<td>1.17113.023</td>
			<td></td>
		</tr>
		<tr>
			<td>Bouin&#39;s fixed solution</td>
			<td>Kaixiu company (Guangzhou city, China)</td>
			<td>20180627</td>
			<td>Sealing</td>
		</tr>
		<tr>
			<td>Dimethylbenzene</td>
			<td>Sinopharm Chemical Reagent limited corporation, China</td>
			<td>20180607</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethyl alcohol (75%)</td>
			<td>Likang company (Nanchang city, China)</td>
			<td>20180915</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethyl alcohol (95%)</td>
			<td>Likang company (Nanchang city,China)</td>
			<td>20181005</td>
			<td></td>
		</tr>
		<tr>
			<td>Glacial acetic acid</td>
			<td>Lingfeng company (Shanghai city, China)</td>
			<td>20170912</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin staining solution</td>
			<td>BASO company (Zhuhai city, China)</td>
			<td>718101</td>
			<td></td>
		</tr>
		<tr>
			<td>Masson kit</td>
			<td>Weigxi company (Guangzhou city, China)</td>
			<td>18121101</td>
			<td>Store at 4&#176;C</td>
		</tr>
		<tr>
			<td>PASM kit</td>
			<td>BASO company (Zhuhai city, China)</td>
			<td>618091</td>
			<td>Store at 4&#176;C</td>
		</tr>
	</tbody>
</table>

modified pasm-masson staining in a renal biopsy

Special histochemical staining is an important method for a renal biopsy, which includes PAS staining, periodic acid silver methenamine (PASM) staining, and Masson staining. In this study, we aim to develop and modify the method of PASM and Masson double staining (PASM-Masson) in a renal biopsy. One hundred and fifty-two cases of renal biopsy were detected by PASM-Masson staining in our hospital. The results showed that the background of the slices was clean, and the renal structures were more visible under the microscope. The glomerular basement membrane, renal tubular basement membrane, and capillary basement membrane were stained black or brown; the mesangium and collagen fibers were stained blue black; the renal tubular epithelial cells and immune complexes were dyed red. These findings indicate that the glomerular immune complex deposition can be accurately identified and positioned by modified PASM-Masson staining, which might have important effects on the pathological diagnosis of renal diseases.

PASM-Masson staining before modification (Figure 1A-D) showed that the hexamine silver staining was dyed deep black, and Masson staining showed a light dyeing on the immune complex in the glomerulus. In addition, the contrast between the basement membrane and the immune complex was not distinct. As a whole, the structures of the glomeruli and renal tubules were indistinct, and the background colors displayed hyperchromasia.
<p class="jove...

In this study, we present a protocol of PASM-Masson dual staining to identify the location and extent of glomerular immune complex deposition in renal diseases.

Modified PASM-Masson Staining in a Renal Biopsy

Special histochemical staining is an important method for a renal biopsy, which includes PAS staining, periodic acid silver methenamine (PASM) staining, ...