How to choose fluorescent dyes in laser confocal

Fluorescent dye selection guide

With the development of life science , A large number of fluorescent dyes play an important role in cell imaging and flow cytometry analysis .

Fluorescent dyes absorb light waves of one wavelength and emit light waves of another wavelength , The wavelengths of absorbed and emitted light constitute the excitation and emission spectra of fluorescent dyes （ It is called fluorescent dye spectrum for short ）.

All instruments that need fluorescent dyes will be equipped with lasers , Lasers produce light of a certain wavelength , So as to excite the fluorescent dye that can absorb the light of this wavelength , Fluorescent dyes are excited , The emitted light is recognized by the instrument after passing through the filter , It is worth noting that each channel can only detect one fluorescent signal within a certain range .

There are a variety of fluorescent dyes in various spectral ranges , How should we choose these fluorescent dyes in the experiment ？

1. Choose fluorescent dyes according to the instrument ：
   1. Only one fluorescent dye can be selected for each channel , And the wavelength is within the wavelength range of the laser ;
   2. The emitted spectrum must be within the appropriate range that the filter of the instrument can accept ;
2. In multicolor experiments that require two or more fluorescent dyes , The fluorescent dyes between the channels can be matched arbitrarily , But spectral overlap should be minimized ;
3. For antigens with low expression, fluorescent dyes with high signal-to-noise ratio should be selected , Such as phycoerythrin （R-Phycoerythrin, R-PE）;
4. When detecting cells with high autofluorescence , Choose fluorescent dyes with longer emission wavelength , Such as Cy Series dyes , Better signal value can be obtained .

The optical characteristics of common fluorescent dyes are as follows ：

notes ：1、 Blue （350-450nm Local excitation ） 2、 green （488nm Local excitation ） 3、 Orange yellow （543-555nm Local excitation ） 4、 Red （568-594nm Local excitation ） 5、 Far infrared （620-660nm Local excitation ） 6、 The near infrared （680-790nm Local excitation ）

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How to choose fluorescent dyes in laser confocal

Dye selector ：

 Fluorescence SpectraViewer