Fungus/Yeast Sample Cell Cell Cycle Iodine Propidium Red Fluorescence Flow Cytometry Kit Instructions

Fungal/Yeast Cell Sample Cell Cycle Propidium Iodide (PROPIDIUM IODIDE) Red Fluorescence

Flow Cytometry Kit Product Manual (Chinese version)

The main purpose

YIJI fungal/yeast cell sample cell cycle propidium iodide (PROPIDIUM IODIDE) red fluorescent flow cytometry reagent is a kind of purpose to quantify the degree of DNA binding propidium iodide dye in fungal/yeast cells by cell flowmeter. DNA content, an authoritative and classical technical approach to further gaining cell cycle distribution. The technology has been carefully developed and successfully tested. It is suitable for cell cycle analysis of various fungal/yeast cells, including budding yeast, fission yeast, and the like. It can be used for research on apoptosis, signal transmission, cell division, metabolism, and the like. The product is strictly sterile, ready to use, high resolution, simple operation and stable performance.

technical background

Saccharomyces cerevisiae and Schizosacchromyces pombe are model organisms that study the cell cycle of eukaryotes. Cell cycle is an important indicator for evaluating cellular metabolic, physiological and pathological conditions. The DNA content is determined by DNA-binding dyes by means of a cell flow meter, and the different stages of the cell cycle are determined and distinguished according to changes in the DNA content as the cell cycle changes. Flow cytometry (FLOW CYTOMETRY) is a high-tech science developed in the 1970s that combines computer, laser, fluid mechanics, cytochemistry, and cellular immunology. Apoptotic cells are subdiploid (sub-G1) cell populations that appear in front of the normal G0/G1 peak cell population.

product content

YIJI cleaning solution (Reagent A) 200 ml

YIJI Fixative (Reagent B) 100ml

YIJI anti-interference solution (Reagent C) 25 ml

YIJI Enzyme Solution (Reagent D) 10 ml

YIJI staining solution (Reagent E) 25 ml

Product manual 1 copy

preservation method

Store YIJI anti-interference solution (Reagent C), YIJI hydrolysate (Reagent D) and YIJI stain solution (Reagent E) in a refrigerator at -20 °C, and keep the rest in a 4 °C refrigerator; YIJI staining solution (Reagent E) to avoid Illumination, effective guarantee for June

User-supplied

2 ml centrifuge tube: container for cell staining

(Micro) Benchtop Centrifuge: For sample handling operations

Incubator or thermostatic bath: for reaction incubation

Cell flow meter: for cell fluorescence analysis

Experimental procedure

• Prepare 5 X 10 ⁶ test fungus/yeast cells
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the supernatant
• Add 1 ml of YIJI cleaning solution (Reagent A) to a 2 ml centrifuge tube and mix.
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the supernatant
• Slowly add 2 ml of pre-cooled YIJI fixative (Reagent B) and mix by vortexing
• Incubate in a refrigerator at 4 ° C for 16 hours
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the fixing solution
• Add 1 ml of YIJI cleaning solution (Reagent A) and mix.
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the supernatant
• Repeat experiment steps 11 through 13
• Add 500 μl of YIJI Anti -Interference Solution (Reagent C ) and mix
• Put in a 37 ° C constant temperature bath and incubate for 2 to 16 hours
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the supernatant
• Add 200 μl of YIJI Enzyme Solution (Reagent D ) and mix
• Put in a 37 ° C constant temperature bath and incubate for 20 minutes
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the supernatant
• Add 500 μl of YIJI Cleaner (Reagent A) and mix
• Place in a micro-tabletop centrifuge for 1 minute at a speed of 16000g
• Carefully remove the supernatant
• Add 500 μl of YIJI staining solution (Reagent E) to mix the cell population
• Ultrasonic treatment of 100 watts, 30% power, 15 seconds
• Incubate for 30 minutes in a 37 ° C incubator in a dark room
• Moved to the cell flowmeter dedicated test tube (can be placed in the 4 ° C refrigerator to be tested)
• Immediate cell flow analysis: red fluorescence (or 488 nm excitation wavelength, 610 nm emission wavelength) appeared at 488 nm argon ion laser, and more than 50,000 cells were observed.

Precautions

• This product is 50 operations
• Wear gloves when handling
• Light must be avoided when incubating
• It is recommended that cell flow analysis be performed immediately after cell staining is completed. Do not put it in the refrigerator at 4 °C for more than 1 week.
• Before the cell flow analyzer is analyzed, the contents must be mixed or sonicated to avoid cell agglomeration.
• The amount of cells detected should not be less than 10,000
• Tell the flow meter technician about the cell type
• Use unstained cells for FSC and SSC settings, then adjust stained cells to unstained cells, including PMT for FL1, FL2, FL3
• Recommended setting:

Detector FSC E00 Gain: 3

Detector FL2-A Voltage: 890 Gain: 2

Flow rate: 500 / sec

• The company provides a series of cell cycle analysis reagent products

Quality Standard

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

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Chengdu JRT Meter Technology Co., Ltd , http://www.rangingsensor.com