Hematological test v4 1/Apr/2023 RIKENMPP_003b_004 test 0 1 testSt test modification

Mouse blood is fractionated and the number of erythrocytes, the number of white blood cells (leucocytes), hematocrit values, hemoglobin content, and the white blood cell differential are measured.

These procedures are applicable to the mouse phenotyping platform at the RIKEN BRC. The tests should be performed by trained staff members who understand the safety and hygiene rules required by RIKEN. Any queries, comments, or suggestions, relating either to this SOP in general or to a specific problem encountered during a procedure, should be addressed to the Blood Tests Unit Leader. Any deviation from this protocol must be reported to the Blood Tests Unit Leader.

Testing is carried out in accordance with common animal handling procedures. Coveralls (in the rearing facility) or laboratory coats (in the laboratory facility) must be worn with gloves at all times in the work area. If animals are dropped on the floor, treat them in accordance with the safety and hygiene guidelines of the rearing facility at RIKEN. If something unexpected happens during the primary screening, it must be quickly reported to the Blood Tests Unit Leader and recorded.

Be certain to add EDTA, or the blood will coagulate within 30 minutes. Also, tap the tube a few times before the analysis to check whether the blood has coagulated at all. Looking through the sample tube under a light facilitates the detection of coagulated blood. Coagulated samples will clog the lines of the automated hematology analyzer, making the analysis impossible without washing the apparatus. The blood would then need to be sampled again. Time from blood sampling to the analysis After the blood sample is taken, hemocytes tend to increase in volume over time. Therefore, the samples should be analyzed within a few hours (up to 5 hours). Ideally, the analysis should be performed immediately after blood sampling. If this is impossible, however, be sure to keep the period of time from blood sampling to the analysis constant.

Use the manual for the automated hematology analyzer (XN-1000V) to ensure quality control.

Automated hematology analyzer (XN-1000V; sysmex)

Microtubes (1.5 mL) Reagents for the XN-1000V system (see the manufacturer's manual)

Use the manual for the automated hematology analyzer (XN-1000V) when analyzing the blood samples.