ExperimentTypeSPT

released
   March 28th, 2019 at 3:16pm

Overview


Experiment Category 
Microscopy
Assay Classification 
Fluorescence Localization
Experimental Purpose 
Single Particle Tracking
Raw Files Available 
Reads (fastq) provided by lab

Assay Description

SPT

Single particle tracking (SPT) is a method to track the position of a single molecule over time in live cells. Advances in microscopy and molecular biology techniques have enabled a broad range of applications.

The protocol involves preparing the target molecule to be identifiable under the microscope, tracking its position over time and analyzing its trajectory. Generally, the preparation of the molecule of interest is performed by genetically modifying it with tags followed by the binding of reporter molecules such as fluorescent dyes. However, depending on the molecule of interest, the goal of the experiment and the type of microscope used, different techniques are also implemented. Following the preparation of the molecule, a high-resolution microscope is used to take snapshots of the position of the molecule at different intervals of time. Tracking a single molecule in a live cell is a demanding task, requiring both sensitivity and accuracy. Tailored to the application, this can be achieved by specialized microscopy techniques like TIRF or PALM. Analysis of the resulting images results in a file that describes the trajectory of the molecule.

4DN processed files are in a format that includes the ID of a trajectory, the time interval, and the coordinates of the position. Further analysis of the trajectory of the molecule is necessary to gain biological insight.