Abstract
As there are significant variations of cell elasticity among individual cells, measuring the elasticity of batch cells is required for obtaining statistical results of cell elasticity. At present, the micropipette aspiration (MA) technique is the most widely used cell elasticity measurement method. Due to a lack of effective cell storage and delivery methods, the existing manual and robotic MA methods are only capable of measuring a single cell at a time, making the MA of batch cells low efficiency. To address this problem, we developed a robotic MA system capable of storing multiple cells with a feeder micropipette (FM), picking up cells one-by-one to measure their elasticity with a measurement micropipette (MM). This system involved the following key techniques: Maximum permissible tilt angle of MM and FM determination, automated cell adhesion detection and cell adhesion break, and automated cell aspiration. The experimental results demonstrated that our system was able to continuously measure more than 20 cells with a manipulation speed quadrupled in comparison to existing methods. With the batch cell measurement ability, cell elasticity of pig ovum cultured in different environmental conditions was measured to find optimized culturing protocols for oocyte maturation.
Highlights
The mechanical properties of living cells play a key role in cell physiology and pathology [1,2,3,4,5] and can help gain insights in cell structures and functions [6].The oocyte elasticity has especially been found to play a vital role in many physiological processes of animal or cells
The process involved three key techniques: Determination of the maximum permissible tilt angle of the measurement micropipette (MM) and feeder micropipette (FM) to ensure that the MM can insert into the FM, adhesion detection to judge if the target cell is adhered to other cells, and separation of the adhesive cells using calculated aspiration pressure in FM if adhesion occurs
The experimental results showed that the zona pellucida (ZP) Young’s modulus
Summary
The mechanical properties of living cells play a key role in cell physiology and pathology [1,2,3,4,5] and can help gain insights in cell structures and functions [6].The oocyte elasticity has especially been found to play a vital role in many physiological processes of animal or cells. The process involved three key techniques: Determination of the maximum permissible tilt angle of the measurement micropipette (MM) and feeder micropipette (FM) to ensure that the MM can insert into the FM, adhesion detection to judge if the target cell is adhered to other cells, and separation of the adhesive cells using calculated aspiration pressure in FM if adhesion occurs. We measured the Young’s modulus of the porcine oocytes by the new batch method (n = 15) and compared the obtained results with those by traditional MA method (n = 15). We measured the Young’s modulus of the porcine oocytes by the new batch method (n = 15) and compared the obtained results with those 2b.y1.t1r.aMditaixoinmaul MmAPemrmetihssoidbl(enT=il1t5A).nTghleeDexepteerrmimineanttiaolnrefosrulMtsedaesumroinngstMraitcerdopthipaet toteur(MsyMst)emandwaFseeadbeler Mtoiccoronptiinpueottues(lFyMop) erate more than 20 cells one by one, and the average manipulation speed for each cell wToasinqsueratdtrhuepmleedasiunricnogmmpiacroispoinpettotee(xMisMtin) gintmoeththeofdeesd(e2r cmelilc/rmopinipvetste0(.5FMce)lbl/ymainc)er(tnain= d2i2s)t.anWce tmoehaosuldreMtdhicretomhdaecehYilnievos ue20rn1e9gd, ’1s0c, xemllo,dtuhleutsiposf othfetwporcminiceropoicpyetettsescualrteurreedquinireddiffteorebnet beansviicraolnlymhe4onofrt1isz obnytathlliys mneowunbtaetdchramtheetrhtohdan(nh=av5e2t)o.
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