Thomas Adams, Curtis Hauck, Cody Gregg, Trent Ruiz, Andrew Lockwood, Maximillian Seligman, and Dr. Brian Rasnow
Abstract
We integrated an inverted digital fluorescence microscope (Etaluma LumascopeTM), a laboratory robot (KiNEDx KX-300), and a focus motor to create an inexpensive, high throughput, screening platform. High throughput biology or biochemistry is generally performed using multi-well microtiter plates. For example, a stack of ten 384-well plates is equivalent to 3,840 test-tubes, or individual assay containers. Our automated microscopy platform takes sequential multi-color images of each well in a stack of plates, without any human intervention. Commercially available lab equipment that performs similar automated microscopy in microtiter plates costs more than $100,000. Our apparatus achieves these capabilities at a fraction of that cost, making automated high throughput microscopy available to more academic and budget-constrained labs.
Unique to our approach is using one robot to move microtiter plates from stack(s) to the microscope stage, and sequentially positioning each well over the microscope for imaging. This eliminates the need for an expensive automated stage. A stepper motor driven belt around the microscope’s focus knob enabled an Arduino microcontroller to automate focus. Manual focus is facilitated via a toggle switch attached to the Arduino inputs. From device drivers to assay protocol, software architecture is based on a hierarchy of short MATLAB functions and scripts. Device drivers for the robot, LumascopeTM, and focus motor (Arduino) support two modes. Simulation mode facilitates debugging by driving a 3-D graphical representation of the apparatus that updates as the protocol executes. Turning off the simulation mode sends commands to the hardware, which responds in real-time. The bulk of the source code executes identically in both modes, thus reducing the need for tedious debugging of the platform in real time.
We will demonstrate the automated microscopy platform in operation, characterize its throughput and focus performance, and explain its novel use.
