Ruisai small classroom 1217-156 The way to store FCM data is FCS2.0 (flow cytometry standard), which uses List Mode. Easy to process and analyze, but lack of intuitiveness, data display usually has one-dimensional histogram, two-dimensional point map, contour map, density map, etc.; data can also be used to make a standard curve for quantitative analysis. Q1: What is the meaning of the axis? 2, 2D map Q2: "Gate" and "Regin"? The color in the flow graph is artificially specified to distinguish between different cell populations, regardless of the fluorescent color!! Q5: So smart, the negative is exactly in the 101 division? ? Sharing is a virtue, then share it.
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1, single parameter histogram
The measurement data of each cell single parameter can be organized into a statistical distribution and displayed in a histogram manner. In the figure, the abscissa indicates the value of the relative intensity of the fluorescent signal or the scattered light signal, and the unit is a channel, which may be a line or a log. The ordinate is generally relative to the number of cells / particles! 
In the two-dimensional map, the abscissa/ordinate indicates the value of the relative intensity of the fluorescent signal or the scattered light signal, which may be a line or a log. A two-parameter map separates the sample cell population to facilitate analysis of cells of interest. 
Gate setting is an important technology in flow cytometry. Only through the optimal door setting method can the data be accurately acquired and analyzed, thus obtaining valuable information in clinical diagnosis and research.
On-line gating(threshold gating) monitoring/acquiring data
Off-line gating: Analyze experimental data. Scattered light gate/fluorescent gate. Scattered light/fluorescence joint gate
Multiple logic gates (combination gates) multiple logic gates G3=R1 and R2 (G3=R1×R2)
Other: G=R1 or R2 G=R1 not R2Q3: Color and fluorescent color in the flow chart? 
Q4: How is the Yin/Yang (N/P) in the flow graph divided?
How to make the M1 negative limit? In fact, the division of this M1 is based entirely on the negative (same type) comparison! As shown in the figure below: The left picture shows the isotype control, that is, the basal expression level of the substance you tested in the negative group can be understood as background, resting state, basal level, unstimulated, etc. The right picture is the positive group, that is, after receiving the stimulation, the functional state, the drug action, and the like. 
In fact, we can artificially adjust the voltage and magnification to the corresponding size, so that the negative right border is at a certain value (usually drawn near 101). Of course, you can also adjust to 102,103 power.
Q6: Why do you need fluorescence compensation?
In flow cytometry, more than two fluorescently labeled antibodies are often used for staining, and there is a certain overlap between the emission spectra of various fluorescent dyes currently used.
The term "fluorescence compensation" refers to the process of correcting fluorescence leakage, such as removing any fluorescent signal from the detector other than the matched fluorescence.
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