Development of a system for simultaneous multiplane image acquisition for live cell

Lead Research Organisation: University of Kent
Department Name: Sch of Biosciences

Abstract

During this project the student will develop a system to allow truly synchronous multi-plane image acquisition, which will significantly improve the time-resolution during 3d-live-cell imaging. Cairn have achieved successful simultaneous imaging at two different depths into a sample. The student will develop and extend this technique using Cairn's novel Multisplit device, which allows an image to be split into 4 equivalent images on the same camera chip. The student will integrate lenses into this apparatus to obtain different focal lengths to image 4 discrete z-planes on the chip. They will then develop this method and use a combination of piezo and light path splitters to optimize control and stability of spatial distribution between images to permit z distances to be defined by the user, according to experimental needs.
The aims for the student are to:
1. Develop a simultaneous 4-plane image acquisition system with the aim of making the method widely available.
2. Make a detailed high temporal resolution multi-dimension study of rapid movements of myosins and dynamic rearrangements of the actin cytoskeleton to further our understanding of their regulation and function

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M015130/1 01/10/2015 31/03/2020
1666608 Studentship BB/M015130/1 01/10/2015 31/03/2020
 
Description 1. Fluorescence imaging optimisation determined microscopy conditions to reduce the damaging effects of light energy on live cell samples.

2. Characterised regulatory sites in the myosin V motor protein, responsible for the transport of materials within living cells. The regulatory sites impact myosin V motility, localisation and its role in cytoskeletal assembly and cell growth cycle.

3. Characterised regulatory a site in the myosin I motor protein, responsible for the uptake of matter by living cells and cell cycle regulation. The regulatory site impacts associated proteins binding to myosin I and contributes to cell morphology maintenance and cell cycle regulation.

Data gathered throughout the PhD was gathered via fluorescence imaging and genetic cloning techniques.
Exploitation Route 1. Light energy damage in live cells during fluorescence imaging may be reduced following the methodology used.

2-3. Data gathered about myosin I and myosin V may be used to further understand the multiple dynamic processes that occur within the cell as well as mutations that cause disruptions to normal function.
Sectors Other