University of California

Pioneer Hi-bred International, Inc.

Michelle A Digman

Ning Ma

Abraham Phillip Lee

Enrico Gratton

Xuan Li

Hongtao Chen

Yue Yun

Mohammad Aghaamoo

Dohyun Lee

Gopakumar Kamalakshakurup

Chiara Stringari

Peter Donovan

Luca Lanzano

Gisela Lin

Xuhao Luo

Marzieh Ataei

Francesco Palomba

A system for determining the viability of an embryo comprises an imaging device, an excitation device configured to direct an excitation energy at an embryo, a controller communicatively connected to the imaging device and the excitation device, configured to drive the excitation device and collect images from the imaging device at an imaging frequency, a processor performing steps comprising acquiring a set of images from the imaging device, performing a Fourier Transformation to generate a set of phasor coordinates, computing a D-trajectory, computing a set of values of additional parameters, comparing the set of values to a set of stored values related to embryos of known viability, and calculating a viability index factor of the embryo from the set of values and the set of stored values. Methods of calculating embryo viability and determining one or more properties of a tissue are also described.

System and method for tissue viability screening

Methods and devices for single cell analysis using fluorescence lifetime imaging microscopy (FLIM) are disclosed. The methods utilize microfluidic devices which use traps to immobilize cells for FLIM analysis. The analysed cells may be sorted before or after imaging and may be plant, animal, or bacterial cells. Analysis of the FLIM data may use a phasor plot and may be used to identify a metabolic pattern of the single cells.

Microfluidic label-free isolation and identification of cells using fluorescence lifetime imaging (FLIM)

A passive, hydrodynamic technique implemented using a microfluidic device to perform co-encapsulation of samples in droplets and sorting of said droplets is described herein. The hydrodynamic technique utilizes laminar flows and high shear liquid-liquid interfaces at a microfluidic junction to encapsulate samples in the droplets. A sorting mechanism is implemented to separate sample droplets from empty droplets. This technique can achieve a one-one-one encapsulation efficiency of about 80% and can significantly improve the droplet sequencing and related applications in single cell genomics and proteomics.

High-efficiency particle encapsulation in droplets with particle spacing and downstream droplet sorting

A label-free imaging method to monitor stem cell metabolism discriminates different states of stem cell as they differentiate in a living tissues. We use intrinsic fluorescence biomarkers and the phasor approach to Fluorescence Lifetime Imaging Microscopy (FLIM). We identify and map intrinsic fluorophores such as collagen, retinol, retinoic acid, flavins, nicotinamide adenine dinucleotide (NADH) and porphyrin. We measure the phasor values of germ cells ingerm line. Their metabolic fingerprint cluster according to their differentiation state, reflecting changes in FAD concentration and NADH binding during the differentiation pathway. The phasor approach to lifetime imaging provides a label-free, fit-free and sensitive method to identify different metabolic state of cells during differentiation, to sense small changes in the redox state of cells and may identify symmetric and asymmetric divisions and predict cell fate.

Phasor method to fluorescence lifetime microscopy to discriminate metabolic state of cells in living tissue

An optical imaging method based on a feedback principle in which the specific scan pattern is adapted according to the shape of the sample. The feedback approach produces nanometer-resolved three dimensional images of very small and moving features in live cells and in a matter of seconds. Images of microvilli in live cultured opossum kidney cells expressing NaPi co-transporter proteins with different GFP constructs and images of cell protrusions in a collagen matrix are produced with a resolution of about 20 nm. Along cell protrusions in three dimensional cellular adhesions could be identified to the extracellular matrix.

Nanometer-scale optical imaging by the modulation tracking (MT) method

Growing community of inventors

Irvine, CA, United States of America

Michelle A Digman