Blue fluorescent cell nuclear staining analysis kit product manual

YIJI Blue Fluorescent Cell Nuclear Staining Analysis Kit Product Manual (Chinese version)

The main purpose

YIJI Blue Fluorescent Cell Nuclear Staining Analysis Reagent is an authority designed to specifically bind to nuclear DNA by using fluorescent dye DAPI and emit fluorescence detection signals for analysis and observation of nuclear morphology or DNA content as well as double staining or overlapping staining. And the classic technical method. The technology has been carefully developed and successfully tested. It is suitable for observation of various cell nuclei (animal, human, plant, insect, etc.). The product is ready to use, stable performance and clear fluorescence.

technical background

4,6-diamidino-2-phenylindole (DAPI), which is one of the cell DNA stains, specifically interacts with the grooved portion of the DNA double helix Thereby tightly binding to the double strand of DNA. The absorption peak of the fluorescing group produced after the combination is 358 nm and the scattering peak is 461 nm, which is exactly the excitation wavelength of 356 nm of UV (ultraviolet light), making DAPI a commonly used fluorescence detection signal and mainly as double staining or overlapping dyeing. One of the stains.

product content

YIJI cleaning solution (Reagent A) 200 ml

YIJI fixative (Reagent B) 50 ml

YIJI Expansion Fluid (Reagent C) 50ml

YIJI staining solution (Reagent D) 20 ml

YIJI anti-quenching agent (Reagent E) 5 ml

Product manual 1 copy

storage method

Store YIJI fixative (Reagent B), YIJI staining solution (Reagent D) and YIJI anti-quenching agent (Reagent E) in a refrigerator at -20 °C; the rest are stored in a 4 °C refrigerator; YIJI staining solution (Reagent D) and YIJI anti-quenching agent (Reagent E) avoids light; effective guarantee for June

User-supplied

Mini Tabletop Centrifuge: Operation for Suspension Cells

Cover slip: for dyed post-sealing

Fluorescence microscopy: for observation of nuclear DNA staining

Experimental procedure

• Adherent cell staining
• Prepare 1 cell culture 24-well plate of cells to be tested, each well concealed 70%

(Note: slides, slides, 35mm culture dishes, and other culture plates can be used, see note 6 )

• Carefully remove the cell culture fluid
• Carefully add 500 μl of YIJI Cleaner (Reagent A)
• Carefully remove the YIJI cleaning solution (Reagent A)
• Carefully add 500 μl to -20 ° C pre-cooled YIJI fixative (Reagent B) along the wall of the well to cover the surface of the well.
• Incubate for 6 minutes in a 4 ° C freezer
• Carefully remove the YIJI fixative (Reagent B)
• Carefully add 500 μl of YIJI Cleaner (Reagent A) along the wall of the well to cover the surface of the well
• Incubate for 5 minutes at room temperature
• Carefully remove 500 μl of YIJI Cleaner (Reagent A)
• Carefully added 500 microliters YIJI transparent solution (Reagent C), covering the surface of culture wells
• Incubate for 5 minutes at room temperature
• Carefully remove 500 μl of YIJI Transparency Fluid (Reagent C)
• Carefully add 500 μl of YIJI Cleaner (Reagent A) to cover the surface of the well
• Carefully remove 500 μl of YIJI Cleaner (Reagent A) and allow to air dry

(Note: Users can perform antibody fluorescent staining or other fluorescent staining at the beginning of this step )

• Carefully add 200 μl of YIJI staining solution (Reagent D) to cover the surface of the culture well
• Incubate for 10 minutes at room temperature to avoid light
• Carefully remove 200 μl of YIJI stain (Reagent D)
• Carefully add 500 μl of YIJI Cleaner (Reagent A) to cover the surface of the well
• Immediate observation under an inverted fluorescence microscope: excitation wavelength 350 nm, emission wavelength 460 nm (nuclear blue)

Suspension cell staining

• Transfer suspended cells (1 X 10 ⁶ cells) into a 1.5 ml centrifuge tube
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove the supernatant
• Carefully add 500 μl of YIJI Cleaner (Reagent A) to mix the cell pellets
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove the YIJI cleaning solution (Reagent A)
• Gently fingering and loosening the cell population
• Add 500 μl of -20 ° C pre-cooled YIJI fixative (Reagent B) one by one to mix the cell pellets
• Incubate for 6 minutes in a 4 ° C freezer
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove the YIJI fixative (Reagent B)
• Carefully add 500 μl of YIJI Cleaner (Reagent A) to mix the cell population
• Incubate for 5 minutes at room temperature
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove 500 μl of YIJI Cleaner (Reagent A)
• Carefully added 500 microliters YIJI transparent solution (Reagent C), mixed cell population of particles
• Incubate for 5 minutes at room temperature
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove 500 μl of YIJI Transparency Fluid (Reagent C)
• Carefully add 500 μl of YIJI Cleaner (Reagent A) to mix the cell population
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove 500 μl of YIJI Cleaner (Reagent A)

(Note: Users can perform antibody fluorescent staining or other fluorescent staining at the beginning of this step )

• Carefully add 200 μl of YIJI staining solution (Reagent D) to mix the cell population
• Incubate for 10 minutes at room temperature to avoid light
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove 200 μl of YIJI stain (Reagent D)
• Carefully add 400 μl of YIJI Cleaner (Reagent A) to mix the cell population
• Place in a mini tabletop centrifuge for 10 minutes at a speed of 300g (or 2000RPM, eg eppendorf 5415)
• Carefully remove 400 μl of YIJI Cleaner (Reagent A)
• Carefully remove 100 μl of YIJI Cleaner (Reagent A) and mix the cell pellets
• Remove 5 μl onto the slide and place the coverslip
• Immediate observation under a fluorescence microscope: excitation wavelength 350 nm, emission wavelength 460 nm (nucleus appears blue)

Slide staining

• Prepare 1 cell slide
• Carefully add 100 μl of YIJI Cleaner (Reagent A)
• Carefully remove the YIJI cleaning solution (Reagent A)
• Carefully add 100 μl -20 ° C pre-cooled YIJI fixative (Reagent B) to cover the surface of the slide
• Incubate for 6 minutes in a 4 ° C freezer
• Carefully remove the YIJI fixative (Reagent B)
• Carefully add 100 μl of YIJI Cleaner (Reagent A) to cover the surface of the slide
• Incubate for 5 minutes at room temperature
• Carefully remove 100 μl of YIJI Cleaner (Reagent A)
• Carefully add 100 microliters YIJI transparent solution (Reagent C), covering the surface of the slide
• Incubate for 5 minutes at room temperature
• Carefully remove 100 μl of YIJI Transparency Fluid (Reagent C)
• Carefully add 100 μl of YIJI Cleaner (Reagent A) to cover the surface of the slide
• Carefully remove 100 μl of YIJI Cleaner (Reagent A) and let it dry in the air.
• Repeat experiment steps 13 through 14

(Note: Users can perform antibody fluorescent staining or other fluorescent staining at the beginning of this step )

• Carefully add 100 μl of YIJI stain (Reagent D) to cover the surface of the slide
• Incubate for 10 minutes at room temperature to avoid light
• Carefully remove 100 μl of YIJI stain (Reagent D)
• Carefully add 100 μl of YIJI Cleaner (Reagent A) to cover the surface of the slide
• Carefully remove 100 μl of YIJI Cleaner (Reagent A)
• Carefully add 50 μl of YIJI anti-quenching solution (Reagent E)
• Cover the coverslip
• Immediate observation under a fluorescence microscope: excitation wavelength 350 nm, emission wavelength 460 nm (nucleus appears blue)

Precautions

• This product is 100 times (4 24-well plates) and 200 (slides) operating specifications
• This product is friendly for double dyeing or overlapping dyeing
• YIJI staining solution (Reagent D) is a semi-permeable dye
• Wear gloves when handling
• Avoid contaminating mother liquor during operation
• This product is suitable for culture cells of various specifications: slides, slide culture dishes, 35mm culture dishes, and various culture plates. The reagent solution should be adjusted accordingly:

• This product can be used to suspend cells or cells after treatment and stain
• The company provides a series of nuclear staining analysis reagent products

Quality Standard

• This product has been certified to be stable.
• This product has been identified as fluorescent

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