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OpenStage: a low-cost motorized microscope stage with sub-micron positioning accuracy.

Campbell RA, Eifert RW, Turner GC - PLoS ONE (2014)

Bottom Line: Home-built multiphoton microscopes are easy to build, highly customizable, and cost effective.We obtain positioning repeatability of the order of 1 μm in X/Y and 0.1 μm in Z.Our "OpenStage" controller is sufficiently flexible that it could be used to drive other devices, such as micro-manipulators, with minimal modifications.

View Article: PubMed Central - PubMed

Affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States of America.

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 1 μm in X/Y and 0.1 μm in Z. A hand-held control-pad allows the user to direct stage motion precisely over a wide range of speeds (10(-1) to 10(2) μm·s(-1)), 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.

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Complete stage setup.This color-coded image shows the main components of our microscope stage. The gantry is constructed out of ThorLabs XT95 rails (yellow). The objective is mounted on a linear translator (red) at the gantry's center. A 24′′ by 24′′ breadboard forms the X/Y stage (1. purple, ThorLabs PBG11105). A raised sub-stage (2. purple, ThorLabs MB1224) brings the specimen up to the level of the objective which is about 13′′ above the surface of the air table. The specimen is mounted on an independently movable platform (cyan), allowing its position to be manipulated manually with respect to the rest of the stage. Our motorization hardware (motors, couplers, and flexible shafts) are colored green.
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pone-0088977-g001: Complete stage setup.This color-coded image shows the main components of our microscope stage. The gantry is constructed out of ThorLabs XT95 rails (yellow). The objective is mounted on a linear translator (red) at the gantry's center. A 24′′ by 24′′ breadboard forms the X/Y stage (1. purple, ThorLabs PBG11105). A raised sub-stage (2. purple, ThorLabs MB1224) brings the specimen up to the level of the objective which is about 13′′ above the surface of the air table. The specimen is mounted on an independently movable platform (cyan), allowing its position to be manipulated manually with respect to the rest of the stage. Our motorization hardware (motors, couplers, and flexible shafts) are colored green.

Mentions: Our 3-axis stage is integrated into our custom-built 2-photon microscope (Fig.1, where key components are color-coded). The microscope is built around a gantry that places the objective over the middle of a lightweight 24′′×24′′ ThorLabs aluminum breadboard. The objective is mounted to a single linear translator, which allows for focusing. Four pairs of linear translators couple the X/Y stage to the air-table and allow it to be moved with respect to the objective (Figs. 2 & 3). Motion in X, Y, and Z is actuated by micrometers (not visible in Fig. 1) coupled to stepper motors via a flexible shaft. The specimen itself sits on a raised sub-stage to enable easy access to it from a variety of angles (including from underneath). The sub-stage is a 12′′ by 24′′ breadboard raised about 10′′ above the base breadboard using 4 posts. Our specimens are mounted on a small custom-machined platform that can be translated in X and Y by two ThorLabs PT1 linear translators (cyan, Fig.1) located on the raised breadboard. This second pair of translators is optional and allows the sample to be accurately positioned with respect to other components on the rig.


OpenStage: a low-cost motorized microscope stage with sub-micron positioning accuracy.

Campbell RA, Eifert RW, Turner GC - PLoS ONE (2014)

Complete stage setup.This color-coded image shows the main components of our microscope stage. The gantry is constructed out of ThorLabs XT95 rails (yellow). The objective is mounted on a linear translator (red) at the gantry's center. A 24′′ by 24′′ breadboard forms the X/Y stage (1. purple, ThorLabs PBG11105). A raised sub-stage (2. purple, ThorLabs MB1224) brings the specimen up to the level of the objective which is about 13′′ above the surface of the air table. The specimen is mounted on an independently movable platform (cyan), allowing its position to be manipulated manually with respect to the rest of the stage. Our motorization hardware (motors, couplers, and flexible shafts) are colored green.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3935852&req=5

pone-0088977-g001: Complete stage setup.This color-coded image shows the main components of our microscope stage. The gantry is constructed out of ThorLabs XT95 rails (yellow). The objective is mounted on a linear translator (red) at the gantry's center. A 24′′ by 24′′ breadboard forms the X/Y stage (1. purple, ThorLabs PBG11105). A raised sub-stage (2. purple, ThorLabs MB1224) brings the specimen up to the level of the objective which is about 13′′ above the surface of the air table. The specimen is mounted on an independently movable platform (cyan), allowing its position to be manipulated manually with respect to the rest of the stage. Our motorization hardware (motors, couplers, and flexible shafts) are colored green.
Mentions: Our 3-axis stage is integrated into our custom-built 2-photon microscope (Fig.1, where key components are color-coded). The microscope is built around a gantry that places the objective over the middle of a lightweight 24′′×24′′ ThorLabs aluminum breadboard. The objective is mounted to a single linear translator, which allows for focusing. Four pairs of linear translators couple the X/Y stage to the air-table and allow it to be moved with respect to the objective (Figs. 2 & 3). Motion in X, Y, and Z is actuated by micrometers (not visible in Fig. 1) coupled to stepper motors via a flexible shaft. The specimen itself sits on a raised sub-stage to enable easy access to it from a variety of angles (including from underneath). The sub-stage is a 12′′ by 24′′ breadboard raised about 10′′ above the base breadboard using 4 posts. Our specimens are mounted on a small custom-machined platform that can be translated in X and Y by two ThorLabs PT1 linear translators (cyan, Fig.1) located on the raised breadboard. This second pair of translators is optional and allows the sample to be accurately positioned with respect to other components on the rig.

Bottom Line: Home-built multiphoton microscopes are easy to build, highly customizable, and cost effective.We obtain positioning repeatability of the order of 1 μm in X/Y and 0.1 μm in Z.Our "OpenStage" controller is sufficiently flexible that it could be used to drive other devices, such as micro-manipulators, with minimal modifications.

View Article: PubMed Central - PubMed

Affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States of America.

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 1 μm in X/Y and 0.1 μm in Z. A hand-held control-pad allows the user to direct stage motion precisely over a wide range of speeds (10(-1) to 10(2) μm·s(-1)), 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.

Show MeSH