First, the laboratory cryo rack, equipment and hardware facilities must be in good condition, and the reagents and measurement methods must be followed. For the sample absorbance value ( A) If it is higher than the CO value, it is judged as positive, and on the contrary, it is judged as negative. It is prepared before use.

4 Judgment of results and report 4. In my actual work, the author found that patients whose results are near the CO value have poor repeatability and are also likely to cause medical disputes. Therefore, it is necessary to select and order long batches of reagents and ensure storage conditions. In summary, the ELISA detection operation steps are complex, and many factors that may affect the measurement result are distributed in each step of the measurement operation. The substrate catalyzed by HRP is hydrogen peroxide.

Serum samples should be centrifuged sufficiently, otherwise false positive results may be caused by non-specific adsorption of fibrinogen in the micropores. If all kinds of acidic terminating liquids are selected, the blue color will be changed to yellow. The ELISA plate can be placed in a water bath box, and the bottom of the plate should be close to the water surface to quickly balance the temperature.

The wet box should use materials with good heat transfer properties (such as metal, etc. In order to ensure the stability of the results, it is necessary to strictly follow the temperature and time specified in the kit instructions, and the temperature and time cannot be changed at will. The hospital strictly implemented the review system in actual work and achieved good results. Therefore, the establishment of a review system and increased review efforts can ensure the accuracy of the results, such as Re-examine the test results of the 'two-and-a-half' hepatitis B in the 'gray zone' and the rare mode.

This system can also add transmission light imaging and color imaging functions on the basis of 4-color fluorescence channel imaging, a variety of magnification objectives are optional, environmental control system (EC), injector system, etc. The application of transmitted light for cell counting and apoptosis detection and analysis software can divide and count cells without resorting to any dye molecules. Autophagy dysfunction and mitochondrial integrity are related to the pathogenesis of various diseases, including neurodegenerative diseases and cardiovascular diseases.

The instrument based on cytology research can use ImageXpress;Pico automatic cell imaging system combined with CellReporter-XpressTM automatic image analysis software. Evaluation of cell morphology and viability Imaging and analysis methods as an effective tool can provide information on a variety of parameters, including cell viability, characteristics of cells at different stages, cell adhesion and distribution status, cytoskeletal integrity (morphology) and mitochondrial potential Change etc. In order to observe the actin cytoskeleton, the cells were fixed with 4% paraformaldehyde (Sigma) and stained with phalloidin linked to AlexaFluor; 488 (AF488).

Multiple parameters including cell viability, apoptosis, mitochondrial membrane potential, and autophagy are of great significance for the study of the toxicity mechanism of living or fixed cells.Poor water quality in the first-grade effluent ①. Insufficient water supply, improve water supply or replace pretreatment filter element. Replace the high-pressure protection switch. At this time, the full water indicator light will be on.

The cryo rack does not make water ①. ②. ③. Replace the low-voltage protection switch. Short-circuiting may cause the booster pump to idle and damage the pump. The system function is abnormal: the system circuit may be damaged, replace the circuit board. Open the pure bucket ball valve. invited a technician, Master Su, to explain in detail the common troubleshooting of the ultra-pure water machine in the laboratory! 1. The circulation pump may be damaged, and the circulation treatment cannot be performed. The amount of water produced by the system is too small. It may be deposited for too long, and drain it., Ltd

If you look up and then head down cryo rack after the probe is down, it may be because the template amount in the system is too high, it is recommended Reuse after dilution. I. Generally speaking, the fluorescence amplification curve can be divided into three stages: the fluorescence background signal stage, the fluorescence signal exponential amplification stage, and the plateau stage. Common housekeeping genes are beta-actin,

GAPDH, 18SrRNA and so on. 3) Specific primers:

The specific reverse transcription method is to use oligonucleotides containing the complementary sequence of the target RNA as primers. Add 30 ~ 50 mu; l of RNase-free water, stand still for 1min, shake for 30sec, and centrifuge immediately. To each tube, 2 times; qPCRTaqMix 12. 1. With this method, all RNA molecules in the system all serve as cDNA * strand templates, and PCR primers impart the required specificity during the amplification process.5ul to 25ul Mix well, 70 5 minutes at ℃, immediately ice bath 5 * buffer4.

It is particularly suitable for detecting the expression of multiple genes. Derivation of 2- △△ CT method (for details, see Real-time quantitative PCR and 2- △△ CT method for analyzing relative gene expression) Supplement: After DNaseI digests the DNA in the sample RNA, the sample needs to be extracted again with chloroform.5ul1. Take care not to touch the pellet. It can be used to detect whether rare genes are expressed and quantitatively detect the expression level of specific mRNAs from a very small number of cells.. Because Real-TimePCR has higher requirements on the quality of RNA samples, it is necessary to choose a suitable one before formal experiments.

There are two bright bands, 28S and 18S, without DNA band contamination. Centrifuge at 3.0ul dNTP (10mM) 2. 2- △△ CT method is a simple method to analyze the relative changes of gene expression in real-time quantitative PCR experiments. Centrifuge at 14000 rpm for 8 min at 4 ° C, and take the supernatant (about 250ul). Re-examine the amplification conditions or redesign and verify the primers. The key to doing the dye method is to find the appropriate primers and prevent pollution. Observe with a gel UV analyzer. Specific experiments The process is as follows: