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. 2015 Aug 5;16(8):18149–18184. doi: 10.3390/ijms160818149

Table 2.

Comparison of advantages and limitations in the techniques used for cell adhesion studies.

Method Strength Weaknesses References
Polyacylamide-traction Force Microscopy (PA-TFM) Real time observation;
No special and expensive equipment needed for fabrication Inexpensive;
Flexible to chemical and mechanical adjustment;
Adaptable to a large variety of cells		 Needs to record both unstressed and stressed state of the substrate;
Suffers from uncertainties in tracking beads’ position		 [47,48,49,50,51,52,53,54]
Micropatterning (Micropost array/micropillar) Real-time observation;
Force quantification easier and more reliable than PA-TFM;
The micropillar stiffness is manipulated by its geometry;
Gives good precision over surface chemical properties on micrometer scale		 Substrate can alter cell’s behavior;
Requires sophisticated equipment to fabricate;
Needs skilled operator;
Sensitivity of the microposts to the particular cell type needs to be optimized		 [26,55,56,57,58,59,60]
Three Dimensional Traction Force Quantification (3D-TFM) Real-time observation;
Flexible to chemical and mechanical adjustment;
Adaptable to a large variety of cells;
Flexible to chemical and mechanical adjustment;
Adaptable to a large variety of cells		 Needs high-end confocal microscope;
Needs high computational processing;
Needs to record both unstressed and stressed state of the substrate;
Suffers from uncertainties in tracking beads position		 [61,62,63,64,65,66,67,68,69,70]
Wash Assay Simple Not a quantitative data, needs further analysis to obtain quantitative data;
Poor reproducibility; Insensitive		 [71,72,73,74]
Resonance Frequency Real-time observation;
Real-time measurement		 Poor reproducibility [36,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95]
Microfluidics Straightforward construction and operation;
Real-time observation and measurement;
Convenience in size (compatible with cell sizes);
Fast and simple to operate;
Non-invasive to cell		 Low detachment force;
Restricted to short-term adhesion		 Attachment events [96,97,98,99,100,101,102,103,107]; Detachment Events [19,24,107]; [197,198,199,200]
Cytodetachment Real-time observation;
Quick detachment of cell;
Range of force produced is high and applicable to long-term adhesion		 Alignment of probe and cell;
Time-consuming;
Needs highly skilled (experienced) operator;
Operator variable;
Cell damage (hard contact);
Expensive equipment;
Not real-time measurement		 [104,105,106]
Micropipette Aspiration Real-time observation and measurement;
Common lab equipments		 Alignment of probe and cell;
High skilled (experienced) operator;
Operator variable;
Cell damage (hard contact)		 [18,108,109]
SCFS-AFM probe Real-time observation Precise data for short term adhesion studies Alignment of probe and cell require micromanipulator;
Time consuming;
Need skilled operator;
Operator variable;
Cell damage (hard contact);
Expensive equipments;
Not real-time measurement		 [105]; [116,117,118,119,120,121,122,123,124]
SCFS-Biomembrane Probe Real-time observation;
Precise data for short term adhesion studies		 Low maximum force (pN);
Restricted to short term adhesion;
High skilled (experienced) operator;
Operator variable;
Probe variable (fluctuation of probe due to thermal excitation)		 [125,126,127,128]
SCFS-Optical Tweezer Real-time observation;
Precise data for short term adhesion studies;
Compatible with microfluidic device		 Low maximum force (pN);
Restricted to short term adhesion;
High skilled (experienced) operator;
Operator variable;
Cell damage		 [129,130,131,132,133,134,135,136,137,138]
Centrifugation Many analysis can be examined in parallel;
Common lab equipments 		Low maximum force (uncomplete detachment);
Only a single force can be applied per experiment;
Nota real-time analysis		 [139,140,141,142,143,144,145,146,147]
Spinning Disk A range of stresses able to be applied in single experiment;
High stresses		 Not a real-time analysis;
Custom-made apparatuses		 [148,149,150,151,152,153,154,155,156,157,158,159,160]
Flow chamber: Radial flow;
Parallel flow		 Radial flow: Ranges of stresses applicable in single experiment;
Real-time cell detachment observation;
Paralel flow: Simple fabrication;
Straightforward operation;
Real-time cell detachment observation		 Radial flow: Low detachment force;
Restricted to short term adhesion;
Paralel flow: Low detachment force;
Restricted to short term adhesion		 [163,164,165,166,167,168,169,170,171,172,173,174,175]; [176,177,178,179,180,181,182,183,184,185,186,187,188,189,190,191,192,193,194,195]