Essen Instruments, Inc.

Sartorius Bioanalytical Instruments, Inc.

National Research Council of Canada

Other

University of Illinois

Environmental Research Institute of Michigan

Kirk S Schroeder

Bradley Neagle

Daniel Appledorn

Eric William Endsley

Brad Neagle

Keith J Morton

Nevine Holtz

David Rock

Chester S Gardner

Cicely Schramm

Grigory Filonov

Eric Endsley

Richard Kast

Bradley D Neagle

Keith Morton

Disclosed herein are ATP biosensor fusion proteins and their use for assaying ATP levels in cells.

Methods and compositions for live cell analysis of intracellular ATP

Systems and methods are provided for automatically imaging and analyzing cell samples in an incubator. An actuated microscope operates to generate images of samples within wells of a sample container across days, weeks, or months. A plurality of images is generated for each scan of a particular well, and the images within such a scan are used to image and analysis metabolically active cells in the well. Tins analysis includes generating a 'range image' by subtracting the minimum intensity value, across the scan, for each pixel from the maximum intensity value. This range image thus emphasizes cells or portions of cells that exhibit changes in activity over a scan period (e.g., neurons, myocytes, cardiomyocytes) while de-emphasizing regions that exhibit consistently high intensities when images (e.g., regions exhibiting a great deal of autofluorescence unrelated to cell activity).

Live cell visualization and analysis

Systems, methods, and apparatuses of controlling fluid flow are disclosed. An apparatus includes a first microplate having a first open portion and defining one or more first wells therein, a second microplate having a second open portion and defining one or more second wells therein, and a pneumatic lid constructed of styrene ethylene butylene styrene (SEBS). The pneumatic lid extends over the first open portion and the second open portion and includes one or more microfluidic channels that fluidly couple the one or more first wells to the one or more second wells. The pneumatic lid provides an airtight seal over the first microplate and the second microplate.

Methods and apparatus for perfusion and environment control of microplate lab ware

Methods and apparatus for perfusion and environment control of microplate labware

Apparatus and methods to improve the Boyden chamber used in cellular biological measurements, allowing quantitative optical microscopy of biological cells in situ without using fluorescent probes or optical staining. In the preferred embodiment, a thin porous membrane separating top and bottom reservoirs includes an array of precisely positioned micropores pores manufactured using a laser-based photo-machining (ablation) process. The membrane may be composed of polyethylene terephthalate (PET), polycarbonate, polyimide, polyether ether ketone (PEEK) or other appropriate material. The pores formed in the membrane may have diameters in the range of 1 to 15 microns and spaced apart at a distance ranging from 10 to 200 microns. A plurality of upper and lower reservoirs may be provided to form a multi-well plate. The invention finds application in a wide range of potential biological applications where Boyden chamber geometries are currently used including co-culture studies, tissue remodeling studies, cell polarity determinations, endocrine signaling, cell transport, cell permeability, cell invasion and chemotaxis assays.

Methods and apparatus for improving in vitro measurements using boyden chambers

Systems, including apparatus and methods, for performing electrophysiological measurements on membranous samples, including living cells, isolated cell fragments (such as organelles), and/or artificial membranes (such as vesicles). The apparatus may include a high-throughput electrophysiological measurement system, and components thereof. This measurement system may include, among others, (1) a fluidics head for transferring samples and/or other compounds to a perforated measurement substrate, (2) a pressure-regulated plenum system for positioning samples on the substrate and subsequently forming a high-resistance electrical seal, (3) an activation system (such as a computer-controlled pulsed UV illumination module) for activating caged compounds, (4) an electronics head for applying and/or measuring voltage and/or current, and/or (5) a computer-controlled analysis system for collecting and/or analyzing data. The methods may include methods for performing high-throughput electrophysiological measurements on transporters and/or voltage or ligand-gated ion channels, sequentially and/or simultaneously.

High-throughput electrophysiological measurement system

An ultraviolet light source is added to a high-throughput electrophysiogical measurement system to allow for rapid chemical stimulation via caged compound release to a plurality of measurement samples. The integrated electrophysiogical measurement system includes a computer-controlled data collection system, an integrated electronics head for making parallel electrical measurements, and an integrated fluidics head used in part to transfer test compounds into the measurement process. This light source, and associated light coupling to a plurality of test samples, is used in conjunction with the system to make effectuate high-throughput electrical measurements with respect to fast-acting, chemically-activated electrophysiological events. The UV-source modification allows for rapid stimulation and measurement of multiple fast ligand-gated ion channel events in parallel.

System for rapid chemical activation in high-throughput electrophysiological measurements

Standard electrophysiology via the patch clamp technique is a well-developed and powerful tool in both academic and industrial research in the study of ion channels and transporters. Although widely accepted as the gold standard for these types of measurements, the patch clamp technique is considered labor intensive and relatively slow, limited to measuring only one biological sample at a time. This invention describes and demonstrates a device whereby electrophysiological measurements can be made on cells or cell membranes in a manner which allows for multiple measurements to be made in parallel, without direct human intervention, thereby enhancing the cost effectiveness, throughput and general applicability of the technique in fields such as pharmaceutical drug screening.

High-throughput electrophysiological measurement apparatus

Measurement of the fluorescence of a layer of cells, disposed in a well, with supernatant liquid thereabove, is greatly enhanced in sensitivity by illuminating the cell layer with a beam of light incident thereupon at a first angle and detecting fluorescence emitted by the cells with a detector which views the illuminated cells at a second angle, wherein at least one of the first or second angles is oblique to the cell layer. By so controlling the geometry of the system, the contribution to background fluorescence by the supernatant liquid is greatly minimized. Sensitivity of the technique is further enhanced by restricting the portion of the illuminated cell layer which is viewed by the detector. The technique may be applied to an apparatus for rapidly scanning the fluorescence of a plurality of samples in a multiple well plate.

Method and apparatus for quantitative fluorescence measurements

Method and apparatus are disclosed for determining pupillary response parameters, such as latency, time of acceleration of constriction, and speed of constriction. In a form of the disclosed method a signal is derived which is representative of measured pupil size as a function of time after a visible light stimulus. The signal is modified to remove therefrom artifacts due to eye movement. Pupillary response parameters can be obtained from the modified signal. In a preferred embodiment, the modifying of the signal includes fitting curves to different portions of the signal, correcting at least one of the fitted curves, and deriving the modified signal from the curves, as corrected.

Growing community of inventors

Ann Arbor, MI, United States of America

Kirk S Schroeder