6 Small Details For Easy Cell Scratch Test

Wound Healing Test, Also Known As "scratch Test", Is A Widely Established Technology To Study The Migration Of Collective Cells In Vitro. Wound Healing Is A Simple And Economical In Vitro Test Method To Study Cell Migration/tumor Invasion. The Experimental Principle Is That When The Cells Grow Into A Monolayer, A Blank Area Is Artificially Created On The Fused Monolayer Cells, Which Is Called "scratch". The Cells At The Edge Of The Scratch Will Gradually Enter The Blank Area To Make The "scratch" Heal& Nbsp; Living Cell Imaging Can Be Used By Laboratories With Good Conditions To Analyze The Migration Of Keratinocytes Or Fibroblasts In Wound Healing Experiments. In Combination With The Included Software Analysis Algorithm, It Is Possible To Delay Quantifying The Speed Of Gap Surface Closure And Closing Elapsed Time. The Advantage Of Using A Time Delay Microscope To Obtain Data Is That Wound Analysis Can Be Recorded Under Determined And Stable Conditions In The Incubator& Nbsp; In General, The Cell Types Used In Wound Healing Tests Aim To Reconstruct The Regenerative Characteristics Of The Epidermis. Immortalized Cell Lines And Primary Cells Are Often Used As Models In This Regard. The Most Commonly Used Cell Types Are Keratinocytes And Fibroblasts. So What Should We Pay Attention To When Making Scratch Experiments? 1.  Cell Density; Scratch Test Is Generally A Combination Of Several Cells, But The Growth Rate Of Each Cell Will Be Different. Therefore, The Culture Time Needs To Be Adjusted According To The Growth Characteristics Of The Cells. When The Cells Cover The Bottom Of The Hole, The Scratch Effect Will Be Better. It Is Recommended That 100% Of The Cells Be Covered. 2.  When Drawing Lines With The Gun Head, Try To Be Vertical. Take A 6-hole Plate As An Example, One Hole Can Draw 6 Lines 3; After Drawing Lines, You Must Wash Them Twice With PBS To Remove The Cells You Have Drawn; Note That The Culture Method Must Be Changed To A Serum Free Or Low Serum Medium, Because: After Scratch, Cells Are Cultured In A Serum Free Medium, Which Means That Within 24 Hours Of Monitoring, The Reduction Of Scratches Is The Result Of Cell Migration, So The Impact Of Cell Proliferation On Cell Migration Should Be Minimized; However, If The Cells Float In A Large Area After Being Converted To Serum-free Culture, The Concentration Of Serum Should Not Be Higher Than 2%. 5.  (5) Put It Into 37 ℃ 5% CO2 Incubator For Cultivation. Samples Can Be Taken At 0, 10, 12 And 15 Time Points (the Specific Time Depends On The Needs Of The Experiment). 6.  (6) Statistical Method: Use Image J Software To Open The Picture, Grab The Lines Drawn By Yourself, And Calculate The Average Distance Between Cells.