Wild-type (WT), C5aR1−/− and C5aR2−/− mice on a C57BL/6J genetic background (n = 5-15) were administered with recombinant mouse C5a (Sino Biological, China) at a dose of 50 μg kg-1 via intravenous injection (tail vein). After C5a injection, one drop of blood was collected from the tail tip to make a blood smear on a slide at 0, 15, 30 and 60 min. Blood smears were stained using a Microscopy Hemacolor® Rapid Staining of Blood Smear Kit (Merck, Germany). Briefly, blood smears were fixed in Hemacolor® Solution 1 (methanol). The slides were then stained with Hemacolor® Solution 2 (Eosin Y), followed by Hemacolor® Solution 3 (Azur B). The slides were washed with 1 x PBS (pH 7.2) and mounted with dibutylphthalate polystyrene xylene. Using a 20x/0.4 NA objective on an Olympus CX21 microscope, first 400 white blood cells were counted for each slide, and the proportion of PMNs (i.e., cells containing granules that are light violet) was then calculated as previously described (Wu et al., 2020 (link)).
Microscopy hemacolor rapid staining of blood smear kit
Manufactured by Merck Group
Sourced in Germany
The Microscopy Hemacolor® Rapid Staining of Blood Smear Kit is a laboratory equipment product used for the rapid staining of blood smears for microscopic examination. The kit provides a fast and efficient way to prepare and stain blood samples for analysis.
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2 protocols using microscopy hemacolor rapid staining of blood smear kit
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Quantifying Neutrophil Response to C5a
2
Quantifying Neutrophil Response to C5a
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Wild-type (WT), C5aR1−/− and C5aR2−/− mice on a C57BL/6J genetic background (n = 5-15) were administered with recombinant mouse C5a (Sino Biological, China) at a dose of 50 μg kg-1 via intravenous injection (tail vein). After C5a injection, one drop of blood was collected from the tail tip to make a blood smear on a slide at 0, 15, 30 and 60 min. Blood smears were stained using a Microscopy Hemacolor® Rapid Staining of Blood Smear Kit (Merck, Germany). Briefly, blood smears were fixed in Hemacolor® Solution 1 (methanol). The slides were then stained with Hemacolor® Solution 2 (Eosin Y), followed by Hemacolor® Solution 3 (Azur B). The slides were washed with 1 x PBS (pH 7.2) and mounted with dibutylphthalate polystyrene xylene. Using a 20x/0.4 NA objective on an Olympus CX21 microscope, first 400 white blood cells were counted for each slide, and the proportion of PMNs (i.e., cells containing granules that are light violet) was then calculated as previously described (Wu et al., 2020 (link)).
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