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Immunofluorescence Staining Of Suspension Cells

source:Qida organism  views:1474  time:2022-08-24

Miss Sister Asked Us How To Dye Suspension Cells? Qida's Solution Is Coming, Let's Collect It Together With The Students Who Do Suspension Cell Dyeing& Nbsp; Experiment Steps: 1; Collect Cells, And Put Cells With A Total Number Of 1 * 10 ^ 6 Or About 1 * 10 ^ 7 Into A 2ml Centrifuge Tube; 2.  Add 2ml PBS And Gently Overturn It Up And Down. Centrifuge 800g For 5 Minutes. Discard The Supernatant. In Order To Completely Eliminate The Influence Of Serum, It Needs To Be Repeatedly Cleaned For 2-3 Times. 3.  Prepare A Clean Glass Slide And Treat It With Gelatin Or Polylysine, Or Purchase The Treated Glass Slide For Experiment. 4.  Use A Pipette To Absorb An Appropriate Amount Of Suspended Cell Liquid, Drop It On The Slide, And Let The Liquid Spread Freely To Form A Monolayer Structure, And Let The Cell Suspension Dry Freely At Room Temperature. 5.  Use A Special Brush For Immunofluorescence To Draw A Circle At The Edge Of The Dry Liquid Surface To Avoid Liquid Loss During Subsequent Operations. 6.  Add An Appropriate Amount Of 4% Paraformaldehyde Buffer Solution, Fix It At Room Temperature For 30 Minutes, And Rinse It With PBS For 3 Times, 5 Minutes Each Time. 7.  0.4% Triton X100 Was Digested At Room Temperature For 30 Minutes, And PBS Was Rinsed 3 Times For 5 Minutes Each Time. 8.  2% BSA Closed, 37 Degrees, 30 Minutes. 9.  Slightly Shake Off The Sealing Liquid, Do Not Wash It, Add The First Antibody, And Incubate It For 4 Degrees Overnight. 10.  PBS Flush 3 Times, 5 Minutes Each Time. 11.  Add Fluorescent Secondary Antibody To Avoid Light And Incubate It At 37 ℃ For 30 Minutes (remember Not To Dry It). 12.  PBS Flush 3 Times, 5 Minutes Each Time. 13.  Add DAPI And 10ul PBS, And Keep Them Away From Light At Room Temperature For 5 Minutes 14; Add The Sealing Agent Containing Anti Fluorescence Quenching Agent& Nbsp; Observation Of THP-1 Immunofluorescence Staining (Qida Biological, Product No.: CD0122): Observe The Staining Results Under The Fluorescence Microscope Through The Liquid Phase. If The Film Is Clean And There Are No Residual Impurities And Fluorescent Particles, Then Seal The Film With Neutral Gum. The Staining Results Were Observed Under Laser Confocal Microscope. Then Use Special Image Processing Software To Superimpose The Dye Images Under Different Excitation Wavelengths, And You Can Get The Coexistence Image You Want& Nbsp; K-562 Immunofluorescence Staining (Qida Biological, Product No.: CD0115); 2、 Small Details For Suspension Cell Staining: 1 After The Film Is Processed, It Is Better To Let It Dry Naturally Before Sealing, Otherwise The Residual Water Or PBS Will Affect The Color Of The Film After Sealing. 2. The Neutral Gum Should Not Be Too Thick, Or It Will Not Spread Easily After Being Dropped On The Film. Therefore, It Is Better To Buy Fresh Neutral Gum, Which Seems Inexpensive. The Bottle Cap Should Be Tightly Closed After Use, And The Light Proof Paper (tin Foil) Should Be Used For Outsourcing And Light Proof Preservation. 3. Neutral Gum Should Not Be Added Too Much, Just A Small Drop. Since The Neutral Gum Is Very Sticky, Do Not Use A Pipette Or Pipette To Add It. Dip It Slightly With A Yellow Tip (remember To Cut The Tip Of The Tip Flat). It May Not Be A Drop. It Is Better To Drop It On The Target Area You Want To Observe, But There Is No Clear Target Area For Cell Suspension Samples. After The Neutral Gum Is Dripped, Cover The Cover Immediately (remember That The Cover Should Be Clean, Otherwise The Previous Efforts Will Be Wasted), Otherwise The Xylene In The Neutral Gum Will Volatilize Easily And Become Very Dry And Thick. 4. Finally, Remind Again That After The Cover Sheet Is Covered, Never Use Tweezers To Press The Cover Sheet, Otherwise The Consequences Will Be Unimaginable, And The Neutral Glue Will Slowly And Evenly Spread Around The Tissue Sheet Under The Weight Of The Cover Sheet.
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