hot salt water test

In the hot salt test, blood cells were examined using hot saline to check the nuclear dissolution. Do not eat too greasy, high-protein foods the day before, and avoid heavy drinking. The alcohol content in the blood directly affects the test results. After 8 pm on the day before the medical examination, you should fast. To ensure complete nuclear nuclei, the first is full-temperature operation. The second is fast. Third, breaking cells without destroying the subcellular organelles is the most critical step in the preparation of the nucleus. Basic Information Specialist classification: cardiovascular examination classification: blood examination Applicable gender: whether men and women apply fasting: fasting Analysis results: Below normal: Normal value: no Above normal: negative: Generally normal. Positive: Common in leukemia. Tips: Do not eat too greasy, high-protein foods the day before, and avoid heavy drinking. Normal value The nucleus is insoluble and the cell morphology is clear. Clinical significance Abnormal result The test result is positive. In the granulocyte system, most of the cells in the following stages of mesangial cells, the nucleus is dissolved and the degree is obvious (++~+++). The nucleus of the granulocytes is less dissolved or insoluble, and to a lesser extent (+ to ++). The nuclear lysis of promyelocytic cells is somewhere in between. Lymph and monocytes are generally insoluble, occasionally mildly dissolved (+). >10% of the immature cells above (+) were used as diagnostic limits. The coincidence rate of acute granulocyte diagnosis was 78.5%, but the acute mononuclear or lymphocytes did not. Helps identify acute leukemia cell types. However, the immature cells are lysed (+) below <10%, and acute myeloid leukemia cannot be completely ruled out. The people who need to be examined have people with obvious characteristics of anemia, hemorrhage, infection and fever. Positive result may be disease: leukemia precautions Taboo before the test: Do not eat too greasy, high-protein foods the day before the test, to avoid heavy drinking. The alcohol content in the blood directly affects the test results. After 8 pm on the day before the medical examination, you should fast. Requirements for inspection: 1. In order to ensure the complete nuclear nucleus, the first is full-temperature operation. The second is fast. Third, breaking cells without destroying the subcellular organelles is the most critical step in the preparation of the nucleus. Compared with the tissue block, the cultured cells, especially the adherent cultured cells, are more difficult to break when homogenized with a glass homogenizer. Therefore, the cells should be cultured up and down by using a small-capacity glass homogenizer and a well-stitched mortar. 2. Calculate the correct centrifugal speed by centrifugal force g. Different centrifuges can accurately calculate the centrifugal speed based on this. 3. Perform WesternBlot and 2D-gel electrophoresis, and directly add the loading buffer to lyse the nucleus. Inspection process 1, sample processing: (1) Tissue homogenization: Weigh 100~200mg fresh tissue such as liver, brain, heart muscle, etc., rinse with PBS or normal saline, wash blood, filter paper, dry it, cut into pieces with scissors and put into small volume glass homogenizer Inside. Add 1.0 mL of pre-cooled LysisBuffer, add 50 ul of Reagent A, and grind the tissue up and down 20 times in an ice bath at 0 °C; there are unmilled tissues, which can be filtered with double gauze. (2) Cultured cell homogenate: Digest the cells, wash them with PBS, and collect the cells by centrifugation at 800 × g for 5-10 min. count. 5×107 cells were needed for each extraction, 1.0 mL of ice-cold LysisBuffer was resuspended, 50ul of ReagentA was added, the cell suspension was transferred to a small-capacity glass homogenizer, and the cells were ground for 20 to 30 times in an ice bath at 0°C. . 2. Transfer the tissue or cell homogenate to a 1.5 ml centrifuge tube and centrifuge at 700 xg for 5 min at 4 °C. The nucleus was deposited at the bottom of the collection tube and the supernatant was discarded. The pellet was resuspended by adding 0.5 mL of ice-cold LysisBuffer. 3. Add another 0.5 mL MediumBuffer to another new centrifuge tube, pipette the resuspension from the previous step, carefully add it to the centrifuge tube along the tube wall, place it on the MediumBuffer, and centrifuge at 700 xg for 5 min at 4 °C. The supernatant was transferred to a new centrifuge tube and the nuclei were deposited at the bottom of the tube. 4. Discard the supernatant, add 0.5mLLysisBuffer to resuspend the nuclear sink in the nuclear sediment, centrifuge at 1000×g for 10 min, discard the supernatant, and obtain a relatively pure nuclear pellet. 5. Resuspend the nuclear pellet with 50-100 μL of StoreBuffer or a suitable reaction buffer and store immediately or at -70 °C. Not suitable for the crowd 1. Patients who have taken thyroid hormones, steroid hormones, etc., may affect the results of the examination and prohibit patients who have recently taken the drug history. 2, special diseases: patients with hematopoietic function to reduce disease, such as leukemia, various anemia, myelodysplastic syndrome, etc., unless the examination is essential, try to draw less blood. Adverse reactions and risks 1, subcutaneous hemorrhage: due to pressing time less than 5 minutes or blood draw technology is not enough, etc. can cause subcutaneous bleeding. 2, discomfort: the puncture site may appear pain, swelling, tenderness, subcutaneous ecchymosis visible to the naked eye. 3, dizzy or fainting: in the blood draw, due to emotional overstress, fear, reflex caused by vagus nerve excitement, blood pressure decreased, etc. caused by insufficient blood supply to the brain caused by fainting or dizziness. 4. Risk of infection: If you use an unclean needle, you may be at risk of infection.