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System for identifying and targeting individual cells within a heterogeneous population for selective extraction of cellular content

A technology targeting cells and target cells, applied in laboratory containers, microbial dissolution, individual particle analysis, etc., can solve problems such as downstream analysis limitations

Pending Publication Date: 2020-01-31
THE GOVERNING COUNCIL OF THE UNIV OF TORONTO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, downstream analysis may be limited due to dilution of cell lysate in microchannels

Method used

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  • System for identifying and targeting individual cells within a heterogeneous population for selective extraction of cellular content
  • System for identifying and targeting individual cells within a heterogeneous population for selective extraction of cellular content
  • System for identifying and targeting individual cells within a heterogeneous population for selective extraction of cellular content

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Embodiment

[0294] Prenatal Genetic Diagnostic Testing Using DMF / Laser-Microbeam Lysis

[0295] The systems and methods of the present disclosure are used with trophoblast cells from cervical mucosa samples (CMS). Trophoblast cells adhered and embedded in thick mucus are not suitable for flow cytometry or microchannels.

[0296] CMS samples were loaded onto DMF devices, seeded and incubated therein. Samples are stained with markers to identify fetal cells. After incubation, the medium is changed, cells are laser lysed, and the lysate is collected for off-chip whole-genome amplification and genotyping. Such as Figure 10 In B, the CVS control sample (upper panel) and matched laser-lysed fetal cell sample (lower panel) show the same heterozygous allelic pattern by QF-PCR. This is also positive for trisomy 21 as indicated by the circled allele. Figure 10 C shows the genetic analysis of the same laser-lysed fetal cell sample analyzed using next-generation sequencing and which showed t...

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Abstract

The present disclosure provides a system and method for identifying and targeting individual cells within a cell population for selective extraction of cellular content and a digital microfluidic device having at least one hydrophilic site for receiving cells, an imaging system including a stage for receiving the digital microfluidic device and an imaging module for identifying at least one targeted cell among the cells at the at least one hydrophilic site. The system includes a pulsed laser source for laser lysing the targeted cell thereby releasing the cell content to produce a lysate. A control system controls the pulsed laser source, the imaging system and the digital microfluidic device and is programmed for coordinating steps of i) movement of droplets on the digital microfluidic device, ii) selection of the at least one targeted cell to be lysed located at the at least one hydrophilic site, iii) illumination of the at least one selected targeted cell by the pulsed laser source to lyse the at least one selected targeted cell to produce lysate, and iv) collection of the lysate.

Description

[0001] This application was submitted on May 6, 2019. The title of the invention is "a system for identifying and targeting individual cells in a heterogeneous population to selectively extract cell contents", and the application number is a divisional case of a patent application with the application number 201780068412.0 Apply. technical field [0002] A device for identifying and targeting individual cells in a heterogeneous cellular environment for lysis that is subjected to further downstream analysis. Methods of using these devices as well as systems and kits comprising these devices are also provided. These devices, systems, methods and kits can be used in a variety of different applications, including testing and diagnostics. Background technique [0003] Over the past decade, it has become clear that populations of cells thought to be identical can in fact be very heterogeneous in nature. Two examples of this are (a) the population of cancer cells that make up a t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/10G01N15/14
CPCG01N15/10G01N15/1468G01N2015/1006G01N2015/149G01N1/28G01N1/286G01N1/40G01N15/00G01N35/00B01L3/502761B01L3/502792B01L7/52B01L2200/0668B01L2300/0867B01L2300/089B01L2400/0421B01L2400/0442C12N1/066C12M1/34C12M1/42C12M1/33G01N33/5005B01L3/502715B01L3/502753B01L2200/0647C12Q1/6806C12Q1/6881C12Q2565/629G01N2015/1081
Inventor A·R·惠勒M·D·张伯伦J·L·拉曼纳M·D·M·德莱顿E·科洛米耶兹D·智泰雅
Owner THE GOVERNING COUNCIL OF THE UNIV OF TORONTO