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Related Experiment Video

Updated: Mar 3, 2026

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Published on: December 22, 2011

Time-Lapse Into Immunofluorescence Imaging Using a Gridded Dish.

Nick Lang¹, Catherine G Chu¹, Andrew D Stephens^1,2

¹Biology, University of Massachusetts Amherst, Amherst, MA, USA.

|March 2, 2026

Summary

This summary is machine-generated.

Time-lapse into immunofluorescence (TL into IF) imaging now directly links live-cell dynamics with static immunofluorescence markers. This simplified method uses gridded dishes, eliminating the need for complex tracking software.

Keywords:

Cell biology dynamics Immunofluorescence Live-cell imaging Mechanobiology Nuclear dynamics Nuclear rupture Static imaging Time-lapse imaging

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Published on: May 14, 2016

Area of Science:

Cell Biology
Mechanobiology
Microscopy Techniques

Background:

Live-cell imaging provides dynamic cellular information, while immunofluorescence offers static molecular details.
Connecting these imaging modalities is crucial for understanding cellular processes like nuclear bleb formation but is often technically challenging.
Existing methods require specialized software or stage position tracking, hindering accessibility.

Purpose of the Study:

To develop a simplified protocol for directly correlating time-lapse live-cell imaging with subsequent immunofluorescence imaging.
To provide an accessible method for cell biologists to link cellular dynamics with molecular composition.
To overcome the limitations of current techniques in connecting live and static imaging.

Main Methods:

Cells were cultured and imaged using time-lapse microscopy on gridded imaging dishes.
The gridded coordinate system on the dish allowed for precise relocation of individual cells after live-cell imaging.
Standard immunofluorescence staining was performed on the same cells, guided by the established coordinates.

Main Results:

The protocol successfully enabled direct matching of cells between time-lapse and immunofluorescence images based on coordinates.
This method eliminated the need for third-party analysis software or complex stage position measurements.
The time-lapse into immunofluorescence (TL into IF) protocol proved straightforward and effective for mechanobiology research.

Conclusions:

Time-lapse into immunofluorescence (TL into IF) imaging provides a direct and accessible link between live-cell dynamics and static immunofluorescence.
The use of gridded imaging dishes is the key innovation, simplifying cell tracking and correlation.
This protocol enhances the ability of cell biologists to study dynamic cellular events and their molecular consequences.