Microscope slides containing mouse thymus sections were xed with ice-cold acetone and stained with recombinant anti-mouse lamin B1 antibody (clone EPR22165-121; Abcam) consisting of 10% HiFCS and 0.2% triton-X 100 in PBS overnight followed by staining with 6.6 µg/ml anti-rabbit IgG conjugated to Alexa Fluor ® 555 (clone H + L, F(ab') 2 ; Cell Signalling Technology). Post wash, tissues were stained with 1 µg/ml DAPI solution (Thermo Fischer ™ ). Images were acquired using an Olympus IX71 inverted uorescence microscope at 10X and 40X magni cation and imaging analysis was performed using Image J software.
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Immunofluorescent Imaging of Mouse Thymus
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Other organizations : University of Birmingham, McMaster University, Institute of Immunology
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Variable analysis
independent variables
- Fixation of mouse thymus sections with ice-cold acetone
- Localization and expression of lamin B1 protein in mouse thymus sections
- Nuclei staining with DAPI
- Incubation of mouse thymus sections with recombinant anti-mouse lamin B1 antibody (clone EPR22165-121; Abcam) consisting of 10% HiFCS and 0.2% triton-X 100 in PBS overnight
- Staining of mouse thymus sections with 6.6 µg/ml anti-rabbit IgG conjugated to Alexa Fluor ® 555 (clone H + L, F(ab') 2 ; Cell Signalling Technology)
- Staining of mouse thymus sections with 1 µg/ml DAPI solution (Thermo Fischer ™ )
- Imaging of mouse thymus sections using an Olympus IX71 inverted uorescence microscope at 10X and 40X magni cation
- Image analysis performed using Image J software
- Positive control: Not specified
- Negative control: Not specified
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