Abstract

Recent progress in intracellular calcium sensors and other fluorophores has promoted the widespread adoption of functional optical imaging in the life sciences. Home-built multiphoton microscopes are easy to build, highly customizable, and cost effective. For many imaging applications a 3-axis motorized stage is critical, but commercially available motorization hardware (motorized translators, controller boxes, etc) are often very expensive. Furthermore, the firmware on commercial motor controllers cannot easily be altered and is not usually designed with a microscope stage in mind. Here we describe an open-source motorization solution that is simple to construct, yet far cheaper and more customizable than commercial offerings. The cost of the controller and motorization hardware are under $1000. Hardware costs are kept low by replacing linear actuators with high quality stepper motors. Electronics are assembled from commonly available hobby components, which are easy to work with. Here we describe assembly of the system and quantify the positioning accuracy of all three axes. We obtain positioning repeatability of the order of in X/Y and in Z. A hand-held control-pad allows the user to direct stage motion precisely over a wide range of speeds ( to ), rapidly store and return to different locations, and execute “jumps” of a fixed size. In addition, the system can be controlled from a PC serial port. Our “OpenStage” controller is sufficiently flexible that it could be used to drive other devices, such as micro-manipulators, with minimal modifications.

Highlights

  • Two-photon microscopy [1,2] has gained great popularity in over the last 20 years for both functional [3,4] and structural [5,6] studies

  • Commercial motorization solutions are expensive and in many cases not designed with microscopy in mind

  • In this paper we present a complete low-cost stage motorization solution comprising a 3-axis drive system and standalone controller unit

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Summary

Introduction

Two-photon microscopy [1,2] has gained great popularity in over the last 20 years for both functional [3,4] and structural [5,6] studies. Investigators can build their own microscope from commonly available components [7] and control the acquisition with open-source software [8,9]. In addition to the microscope itself, a 3-axis motorized stage is important as it enables a range of applications such as time-lapse imaging of multiple locations in a sample, automated tracking, building image mosaics, and acquiring 3-D (volume) data. Whilst cheap linear translator are available, the motorized actuators and the controller units to drive them cost many thousands of dollars. These costs are in addition to the hardware required to build the stage itself. In this paper we present a complete low-cost stage motorization solution comprising a 3-axis drive system and standalone controller unit. We dub our system ‘‘OpenStage’’ in recognition of the open source Arduino microcontroller at its heart

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