Tissues from mice were placed into 10% buffered formalin. Slides were prepared and stained in the CCHMC Pathology Core. GCs were enumerated by immunohistochemistry (IHC) using anti-B220 antibody (1:50, BD Biosciences) and anti-peanut agglutinin (PNA; Vector Labs 1:50). All imaging was performed using an Aperio digital slide scanner at 20x magnification, and images were analyzed using ImageScope software (Leica Biosystems).
Digital slide scanner
Manufactured by Leica
Sourced in United States
The Digital Slide Scanner is a high-performance device designed for the digitization of microscope slides. It captures high-resolution images of specimens, enabling efficient storage, sharing, and analysis of slide-based samples.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using digital slide scanner
1
Quantifying Germinal Center Cells
2
Quantifying Germinal Center Cells
3
Histological Analysis of Perfused Brains
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The animal was euthanised at 3 hours post-infusion and was transcardially perfused with 0.9% saline. The brains were then xed in neutral-buffered formalin before being processed and para n embedded.
Coronal sections of 5 and 10 µm were cut and stained with hematoxylin and eosin. Images were scanned using a digital slide scanner (Aperio Technologies, Vista, CA, USA) and image analysis was carried out within the Aperio programme. Image analysis was con rmed by an independent investigator and any cases with > 10% discrepancy were agged for review.
Coronal sections of 5 and 10 µm were cut and stained with hematoxylin and eosin. Images were scanned using a digital slide scanner (Aperio Technologies, Vista, CA, USA) and image analysis was carried out within the Aperio programme. Image analysis was con rmed by an independent investigator and any cases with > 10% discrepancy were agged for review.
4
Reclassification and Immune Influx Analysis
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Reclassification of cases was performed independently by two experienced neuropathologists (WGSS and WVH). All scoring was done blinded to patient characteristics and results of other staining by three independent observers (JFV, WGSS, WVH). Analysis of the immune influx was performed on whole slides at 20x magnification. Immune influx was corrected for the size of tissue on the slide and the tumor percentage as calculated with the manufacturers algorithm based on digitalized immunochemical slides using a digital slide scanner (Aperio Technologies Inc.). Serpin expression was scored as present (+) or absent (-), because serpins were uniformly expressed throughout the tumor.
5
Quantifying Ischemic Infarct Volume
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Animals were euthanised at 3 h post-infusion and transcardially perfused with 0.9% saline. The brains were then fixed in neutral-buffered formalin before being processed and paraffin embedded. Coronal sections of 5 and 10 μm were cut and stained with haematoxylin and eosin. Images were scanned using a digital slide scanner (Aperio Technologies, Vista, CA, United States) and image analysis was carried out within the Aperio program. Image analysis was confirmed by an independent investigator and any cases with >10% discrepancy were flagged for review. Infarct volume (corrected for oedema) was determined by subtracting the measured interhemispheric volume difference (oedema volume, ipsilateral–contralateral) from the measured infarct volume for each slice.
6
Neuropathological Evaluation of Immune Influx
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Reclassification of cases was performed independently by two experienced neuropathologists (WGMS and WVH). All scoring was done blinded to patient characteristics and results of other staining by three independent observers (JFV, WGMS, WVH). Analysis of the immune influx was performed on whole slides at 20x magnification. Immune influx was corrected for the size of tissue on the slide and the tumor percentage as calculated with the manufacturers algorithm based on digitalized immunochemical slides using a digital slide scanner (Aperio Technologies Inc.). SerpinB1, SerpinB4, SerpinB9, CD1 d, HLA-A and HLA-B expression was scored as present (+) or absent (−), using a cut-off of 5% positivity.
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