Available tissue sections were used for immunohistochemistry (IHC) analyses of these 5 NMC cases. Surgical tissue sections were sectioned at 4 μm thickness and then stained with primary antibodies specific for NUT (ab122649, 1 : 500, Abcam, UK), CK (AE1/AE3, prediluted; Dako, Denmark), vimentin (V9, prediluted; Dako), CK7 (EP16, 1 : 200, ZSGB‐BIO, China), SMA (1A4, 1 : 200, ZSGB-BIO), CD34 (QBEND, prediluted, Dako), p63 (ZM-0406, prediluted, ZSGB‐BIO), p40 (ZA-0483, 1 : 100, ZSGB‐BIO), Syn (DAK-SYNAP, prediluted, Dako), CD56 (123C3, prediluted, Dako), CgA (DAK-A3, prediluted, Dako), and Ki-67 (MIB-1, prediluted, Dako). A Dako Omnis automated staining platform was used for the IHC staining of these formalin-fixed paraffin-embedded whole tissue sections. Appropriate positive controls were used for all assays.
Omnis automated staining platform
Manufactured by Agilent Technologies
The Omnis automated staining platform is a laboratory equipment solution designed for the automated preparation and staining of biological samples. It performs various staining techniques to aid in the visualization and analysis of cellular and tissue samples. The Omnis platform is intended to streamline and standardize the staining process, improving efficiency and consistency in the laboratory.
Automatically generated - may contain errors
Lab products found in correlation
Mib 1, by Agilent Technologies (1 mentions)
Ck ae1 ae3, by Agilent Technologies (1 mentions)
Vimentin v9, by Agilent Technologies (1 mentions)
Dak synap, by Agilent Technologies (1 mentions)
Dak a3, by Agilent Technologies (1 mentions)
P stat3, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Vimentin, by Agilent Technologies (1 mentions)
α sma, by Agilent Technologies (1 mentions)
Ventana dab detection kit, by Roche (1 mentions)
4 protocols using omnis automated staining platform
1
Comprehensive Immunohistochemical Analysis of NMC
2
Immunohistochemical Analysis of Mouse CRC Xenografts
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Mouse CRC xenografts were routinely formalin‐fixed and paraffin‐embedded in the Department of Pathology, Hospital Universitario Puerta de Hierro. Sections of 4 µm were stained with hematoxylin and eosin according to standard protocols or processed for immunohistochemistry using the Dako‐Omnis automated staining platform. Primary antibodies anti‐Ki‐67 (Clone MIB‐1; Dako‐Agilent), α‐SMA (Clone 1A4; Dako‐Agilent), p‐STAT3 (Clone D3A7; Cell Signaling Technology), pAKT (Clone D9E; Cell Signaling Technology), ALDH (Clone 44/ALDH; BD Biosciences), and vimentin (Clone V9; Dako‐Agilent) were developed using the EnVision Flex, Low and High pH visualization system.
3
Immunohistochemical Staining of TFE3
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A Dako Omnis automated staining platform was used for immunohistochemical staining of whole tissue sections (4 μm thick) embedded in paraffin and formalin-fixed. For details regarding the antibodies used in this study, see Table 1 . The antibodies were optimized using Ventana DAB detection kit (Ventana Medical Systems), and standard quality control procedures were carried out.
The immunohistochemical staining of TFE3 expressions was quantitatively scored on a scale of 0 to 3+ depending on the extent and the intensity of staining. The sample scored 0 if it did not stain at all; 1, if it stained weakly (light brown); 2, if it stained moderately intense (brown); and 3 in case of high intensity (dark brown). The percentage of TFE3-positive nuclei was scored as follows: 3 (>70%), 2 (40-70%), 1 (10-40%), and 0 (<10%). A histologic score was then generated by multiplying these two scores together, and the final positivity scores were assigned based on this histological score as follows: 3+ (a score of 9), 2+ (a score of 4 or 6), 1+ (a score of 2 or 3), and negative (a score of 0 or 1).
The immunohistochemical staining of TFE3 expressions was quantitatively scored on a scale of 0 to 3+ depending on the extent and the intensity of staining. The sample scored 0 if it did not stain at all; 1, if it stained weakly (light brown); 2, if it stained moderately intense (brown); and 3 in case of high intensity (dark brown). The percentage of TFE3-positive nuclei was scored as follows: 3 (>70%), 2 (40-70%), 1 (10-40%), and 0 (<10%). A histologic score was then generated by multiplying these two scores together, and the final positivity scores were assigned based on this histological score as follows: 3+ (a score of 9), 2+ (a score of 4 or 6), 1+ (a score of 2 or 3), and negative (a score of 0 or 1).
4
Automated Immunohistochemical Staining Protocol
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The antibodies used for these studies are shown in Table 1 . The antibodies were configured as FLEX Ready-to-Use and used with the EnVision FLEX, High pH visualization system according to the manufacturer’s instructions for use. At each testing laboratory, reagents from 1 lot of reagents was used for staining on 1 Dako Omnis automated staining platform. All antibodies and associated reagents were obtained from Dako (Glostrup, Denmark).
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