Using Microfluidic Devices to Measure Lifespan and Cellular Phenotypes in Single Budding Yeast Cells

Ke Zou; Diana S. Ren; Qi Ou-yang; Hao Li; Jiashun Zheng

doi:10.3791/55412

El uso de dispositivos de microfluidos para Medir Lifespan y Celular fenotipos en células individ...

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Using Microfluidic Devices to Measure Lifespan and Cellular Phenotypes in Single Budding Yeast Cells

El uso de dispositivos de microfluidos para Medir Lifespan y Celular fenotipos en células individuales levadura en ciernes

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09:18 min

March 30, 2017

DOI: 10.3791/55412-v

Ke Zou^1,2, Diana S. Ren², Qi Ou-yang^1,3, Hao Li², Jiashun Zheng²

¹The State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, School of Physics,Peking University, ²Department of Biochemistry and Biophysics,University of California, San Francisco, ³Peking-Tsinghua Center for Life Sciences at Center for Quantitative Biology, Academy for Advanced Interdisciplinary Studies,Peking University

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Overview

This article presents a protocol for creating specialized microfluidic devices to measure the lifespan and cellular phenotypes of single yeast cells. This method aims to address key questions in the yeast aging field, particularly regarding the aging processes of mother cells.

Key Study Components

Area of Science

Microfluidics
Cellular biology
Yeast aging

Background

Microfluidic devices allow for precise control of the cellular environment.
Understanding yeast aging can provide insights into broader biological aging processes.
Tracking mother cells and their molecular markers is crucial for this research.
Proper loading of yeast into microfluidic devices is challenging and requires practice.

Purpose of Study

To measure the lifespan of single yeast cells.
To investigate the molecular events leading to cellular aging.
To correlate lifespan with various molecular markers.

Methods Used

Fabrication of microfluidic devices.
Loading of yeast cells into the devices.
Microscope setup for observation.
Tracking of cellular phenotypes and lifespan.

Main Results

Successful tracking of mother cells and their lifespans.
Identification of molecular markers associated with aging.
Insights into the aging process of yeast cells.
Potential applications in other areas of cellular research.

Conclusions

The microfluidic method provides valuable insights into yeast aging.
Challenges in device fabrication and cell loading can be overcome with practice.
This technique may be applicable to other research areas requiring long-term cellular observation.

Frequently Asked Questions

What is the main advantage of using microfluidic devices?

Microfluidic devices allow for precise tracking of single cells and their lifespans while monitoring various molecular markers simultaneously.

Why is it important to study yeast aging?

Studying yeast aging can provide insights into the fundamental processes of cellular aging, which may be relevant to other organisms.

What challenges do researchers face when using this method?

Researchers may struggle with the fabrication of microfluidic devices and the proper loading of yeast cells, which require significant practice.

Can this method be applied to other types of cells?

Yes, while focused on yeast, the method can be adapted for monitoring other cell types and their responses over generations.

What molecular events are being investigated in this study?

The study investigates early molecular events that lead to the loss of cellular homeostasis and contribute to aging and cell death.

How does this research contribute to the field of cellular biology?

This research enhances understanding of aging mechanisms at the cellular level, potentially informing broader biological aging studies.

Este artículo presenta un protocolo optimizado para la producción de chips de microfluidos y la configuración de los experimentos de microfluidos para medir la vida útil y los fenotipos celulares de células de levadura individuales.

El objetivo general de este experimento es fabricar un dispositivo microfluídico especializado y utilizarlo para medir la vida útil y los fenotipos celulares de una sola célula de levadura. Este método puede ayudar a responder algunas de las preguntas clave en el campo del envejecimiento de la levadura, como por qué las células madre envejecen y mueren y cuáles son los eventos moleculares anteriores que impulsan la pérdida de la homeostasis celular y, finalmente, el envejecimiento y la muerte celular. La principal ventaja de esta técnica es que podemos rastrear las células madre, rastrear su vida útil y los diversos marcadores moleculares al mismo tiempo, por lo que podemos buscar eventos moleculares que se correlacionen con la vida útil.

Aunque este método puede proporcionar información sobre el envejecimiento de la levadura, también se puede aplicar a otras áreas, como el monitoreo de las respuestas secundarias a la participación que requiere el control celular durante muchas generaciones. Por lo general, las personas nuevas en este método tendrán dificultades porque la fabricación de dispositivos microfluídicos, la carga de células y la configuración del microscopio requieren mucha práctica para funcionar con éxito. La demostración previa de este método es fundamental, ya que es difícil cargar correctamente el dispositivo microfluídico con levadura.

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