Livers from age and sex matched mice were frozen in optimal cutting temperature (O.C.T., Sakura). Then 7 μm sections were cut, fixed with acetone for 10 min, and stained with anti-CD68 Alexa Fluor 647 (FA-11, BioLegend). After washing in PBS, nuclei were stained with Hoechst 33258 for 5 min prior to mounting in Prolong Diamond Antifade mounting medium (Invitrogen). Sections were imaged using the Keyence BZ-X710 fluorescence microscope. CD68+% and area was determined using FIJI software54 (link). Briefly, color images were split into channels and a threshold set for each channel. The analyze particles tool was used to estimate number of nuclei in image (Hoechst blue channel). The analyze particle tool was then applied to the CD68+ channel (Red channel). The percentage was then calculated. The measure tool was used to calculate the average area of the particles above the threshold in the CD68+ channel.
Anti cd68 alexa fluor 647 fa 11
Manufactured by BioLegend
Anti-CD68 Alexa Fluor 647 (FA-11) is a fluorochrome-conjugated monoclonal antibody that targets the CD68 antigen, a glycoprotein expressed by monocytes and macrophages. This product is intended for use in flow cytometry and other immunoassay applications.
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3 protocols using anti cd68 alexa fluor 647 fa 11
1
Quantifying CD68+ Macrophages in Mouse Liver Sections
2
Quantifying CD68+ Macrophages in Mouse Liver Sections
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Livers from age and sex matched mice were frozen in optimal cutting temperature (O.C.T., Sakura). Then 7 μm sections were cut, fixed with acetone for 10 min, and stained with anti-CD68 Alexa Fluor 647 (FA-11, BioLegend). After washing in PBS, nuclei were stained with Hoechst 33258 for 5 min prior to mounting in Prolong Diamond Antifade mounting medium (Invitrogen). Sections were imaged using the Keyence BZ-X710 fluorescence microscope. CD68+% and area was determined using FIJI software54 (link). Briefly, color images were split into channels and a threshold set for each channel. The analyze particles tool was used to estimate number of nuclei in image (Hoechst blue channel). The analyze particle tool was then applied to the CD68+ channel (Red channel). The percentage was then calculated. The measure tool was used to calculate the average area of the particles above the threshold in the CD68+ channel.
3
Quantifying Immune Cells in Murine Liver
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Livers from mice 2 or 12 weeks of age and sex matched were frozen in optimal cutting temperature (O.C.T., Sakura). Then 7 μm sections were cut, fixed with acetone for 10 min, and stained with anti-CD68 Alexa Fluor 647 (FA-11, Biolegend). After washing in PBS, nuclei were stained with Hoechst 33258 for 5 min prior to mounting in Prolong Diamond Antifade mounting medium (Invitrogen). Sections were imaged using the Keyence BZ-X710 fluorescence microscope. Images were analyzed using Image J vers 2.0.0-rc-65/1.51n.
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