Hematoxylin and eosin (H&E) staining was performed as previously described and slices were scored by a licensed veterinary pathologist (Data S1 for more information). For lectin staining, using a Dako delimiting pen (Agilent Technologies, Santa Clara, CA), dams were created around tissue sections. The sections were then incubated in 1% BSA (w/v) in approximately 300 μL of PBS for 30 min at RT, then incubated in 10 μg/mL of SNA lectin for 2 h. They were then rinsed for 10 min three times with 1% BSA in PBS. The sections were then incubated in approximately 300 μL of DAPI for 20 min and rinsed with PBS for 5 min. Finally, two drops of Vectashield fluorescence mounting medium (Dako, Agilent Technologies) was added and the sections were cover-slipped and sealed with nail polish. Confocal images were obtained using a fluorescence microscope (Leica Biosystems). Adjacent tissue sections incubated in 1% BSA in PBS served as controls. Zeiss LSM700 confocal microscope connected to a Zeiss Axiovert 200 M with an EC Plan-Neofluar 40x/1.30 numerical aperture oil immersion objective (Carl Zeiss AG, Oberkochen, Germany). Processing of the images was performed using Zen software and applied on the entire images as well as on controls. The presented pictures are representative of independent triplicates.
Vectashield fluorescence mounting medium
Manufactured by Agilent Technologies
Vectashield is a fluorescence mounting medium designed to preserve fluorescent signals in microscopy samples. It is a glycerol-based solution that helps maintain the brightness and integrity of fluorophores, enabling long-term storage and imaging of fluorescently labeled specimens.
Automatically generated - may contain errors
2 protocols using vectashield fluorescence mounting medium
1
Lectin Staining of Tissue Sections
2
Lectin staining and confocal imaging
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HES staining was performed as previously described and slices were scored by a licensed veterinary pathologist (Supplementary Data 1 for more information). For lectin staining, using a Dako delimiting pen (Agilent Technologies, Santa Clara, CA), dams were created around tissue sections. The sections were then incubated in 1% BSA (w/v) in approximately 300 µL of PBS for 30 min at RT, then incubated in 10 µg/mL of SNA lectin for 2 h. They were then rinsed for 10 min three times with 1% BSA in PBS. The sections were then incubated in approximately 300 µL of DAPI for 20 min and rinsed with PBS for 5 min. Finally, two drops of Vectashield fluorescence mounting medium (Dako, Agilent Technologies) was added and the sections were cover-slipped and sealed with nail polish. Confocal images were obtained using a fluorescence microscope (Leica Biosystems). Adjacent tissue sections incubated in 1% BSA in PBS served as controls. Zeiss LSM700 confocal microscope connected to a Zeiss Axiovert 200 M with an EC Plan-Neofluar 40x/1.30 numerical aperture oil immersion objective (Carl Zeiss AG, Oberkochen, Germany). Processing of the images was performed using Zen software and applied on the entire images as well as on controls. The presented pictures are representative of independent triplicates.
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