Studying Cell Rolling Trajectories on Asymmetric Receptor Patterns

作者：Chia-Hua Lee1, Suman Bose2, Krystyn J. Van Vliet1, Jeffrey M. Karp3, Rohit Karnik2 1Department of Materials Science and Engineering,MIT - Massachusetts Institute of Technology, 2Department of Mechanical Engineering,MIT - Massachusetts Institute of Technology, 3HST Center for Biomedical Engineering and Harvard Stem Cell Institute,Brigham and Women's Hospital and Harvard Medical School

简介：We describe a protocol to observe and analyze cell rolling trajectories on asymmetric receptor-patterned substrates. The resulting data are useful for engineering of receptor-patterned substrates for label-free cell separation and analysis.

Overview

This protocol observes and analyzes cell rolling trajectories on asymmetric receptor patterns to facilitate engineering of receptor pattern substrates for label-free cell separation and analysis.

Key Study Components

Area of Science

Cell biology
Biomaterials
Microfluidics

Background

Cell rolling is a critical process in various biological interactions.
Asymmetric receptor patterns can influence cell behavior.
Label-free techniques are essential for cell separation and analysis.
Micro contact printing is a useful method for creating patterned substrates.

Purpose of Study

To develop a protocol for observing cell rolling on receptor-patterned substrates.
To analyze the trajectories of rolling cells for better substrate engineering.
To provide insights into cell movement dynamics under flow conditions.

Methods Used

Micro contact printing of PEG molecules on a gold substrate.
Creation of alternating patterns of self-assembled monolayers.
Backfilling the substrate with P-selectin and receptors.
Flow chamber incorporation and imaging of HL-60 cells rolling on patterns.

Main Results

Image sequences of HL-60 cell interactions were recorded.
Trajectories of rolling cells were analyzed using MATLAB.
Data on edge tracking length and rolling velocities were obtained.
Insights into cell movement on asymmetric receptor patterns were provided.

Conclusions

The protocol enables detailed analysis of cell rolling dynamics.
Asymmetric receptor patterns can be engineered for effective cell separation.
This method contributes to the understanding of cell behavior in shear flow.

Frequently Asked Questions

What is the significance of cell rolling?

Cell rolling is crucial for immune response and tissue repair processes.

How does the protocol improve cell separation?

It allows for label-free separation by utilizing receptor interactions.

What are the advantages of using asymmetric patterns?

Asymmetric patterns can enhance the specificity and efficiency of cell interactions.

What type of cells were used in this study?

HL-60 cells, a human promyelocytic leukemia cell line, were used.

What software was used for data analysis?

MATLAB was utilized for analyzing the rolling cell trajectories.

Can this method be applied to other cell types?

Yes, the protocol can be adapted for various cell types depending on the receptors used.

标签: Cell Rolling Trajectories, Asymmetric Receptor Patterns, Label-free Separation, Analysis Of Cells, Lateral Displacement, Continuous-flow Separation, Receptor Expression, Cell Phenotypes, Adhesion Modulation, Cell Rolling Adhesion, P-selectin Receptors, Microcontact Printing, Gold-coated Slides, Flow Chamber, HL60 Cells, PSGL-1 Ligand, Inclined Edges, Pattern Edges, Edge Tracking Length,