Automated microbeam observation environment for biological analysis—Custom portable environmental control applied to a vertical microbeam system

Matthew J England,Alan W Bigelow,Michael J Merchant,Eirini Velliou,David Welch,David J Brenner,Karen J Kirkby

doi:10.1016/j.snb.2016.08.076

Matthew J England, Alan W Bigelow + Show 5 more

Open Access

https://doi.org/10.1016/j.snb.2016.08.076

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Abstract

Vertical Microbeams (VMB) are used to irradiate individual cells with low MeV energy ions. The irradiation of cells using VMBs requires cells to be removed from an incubator; this can cause physiological changes to cells because of the lower CO2 concentration, temperature and relative humidity outside of the incubator. Consequently, for experiments where cells require irradiation and observation for extended time periods, it is important to provide a controlled environment. The highly customised nature of the microscopes used on VMB systems means that there are no commercially available environmentally controlled microscope systems for VMB systems. The Automated Microbeam Observation Environment for Biological Analysis (AMOEBA) is a highly flexible modular environmental control system used to create incubator conditions on the end of a VMB. The AMOEBA takes advantage of the recent “maker” movement to create an open source control system that can be easily configured by the user to fit their control needs even beyond VMB applications. When applied to the task of controlling cell medium temperature, CO2 concentration and relative humidity on VMBs it creates a stable environment that allows cells to multiply on the end of a VMB over a period of 36h, providing a low-cost (costing less than $2700 to build), customisable alternative to commercial time-lapse microscopy systems. AMOEBA adds the potential of VMBs to explore the long-term effects of radiation on single cells opening up new research areas for VMBs.

Highlights

The environment for in vitro cell culture is typically an incubator at 37 ◦C, 5% CO2 and a relative humidity of 97–100% [1,2,3]
The spikes seen every two hours in the relative humidity, cell dish temperature and ambient CO2 are caused by the removal of the cell dish from the system so that it can be weighed and would not occur if the system was left for normal operation
The Automated Microbeam Observation Environment for Biological Analysis (AMOEBA) was designed to be a modular environmental control system that would be flexible enough to be applied to many situations beyond the area of environmental control

Summary

Introduction

The environment for in vitro cell culture is typically an incubator at 37 ◦C, 5% CO2 and a relative humidity of 97–100% [1,2,3]. Observation of cells with a microscope requires them to be removed from an incubator. This change in environmental conditions experienced by the cells during an experiment is sub-optimal, especially where long-term experiments are desired, to observe factors such as cell motility, mutagenesis or the effects of radiation over several cell cycles. To combat the effects of removing the cells from the incubator, specialised time-lapse microscopes can be purchased that incorporate dedicated environmental control systems. Offthe-shelf time-lapse systems usually fall into two main categories: they are either an incubator that contains a microscope, an example of this type of system is the Nikon BioStation IM-Q (Nikon, Minato, Tokyo, Japan), or they are microscopes with environmental chambers, such as the Cell Observer system by Zeiss (Zeiss, Oberkochen, Stuttgart, Germany) or the Leica DMi8 Live Cell Imaging System (Leica, Wetzlar, Giessen, Germany). Costing up to $150,000, these high-cost systems have driven scientists to design their own timelapse system on a lower budget, an example is the LOCOMOTIS (Low-Cost Motility Tracking System) produced by Lynch et al [5]

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