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

Uso di dispositivi microfluidici misurare la durata della vita e cellulare fenotipi in singole ce...

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

Uso di dispositivi microfluidici misurare la durata della vita e cellulare fenotipi in singole cellule di lievito in erba

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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.

Questo articolo presenta un protocollo ottimizzato per la produzione di chip microfluidici e la configurazione degli esperimenti microfluidici per misurare la durata di vita e fenotipi cellulari delle singole cellule di lievito.

L'obiettivo generale di questo esperimento è quello di realizzare un dispositivo microfluidico specializzato e utilizzarlo per misurare la durata della vita e i fenotipi cellulari di una singola cellula di lievito. Questo metodo può aiutare a rispondere ad alcune delle domande chiave nel campo dell'invecchiamento del lievito, come ad esempio perché le cellule madri invecchiano e muoiono e quali sono gli eventi molecolari precoci che guidano la perdita dell'omeostasi cellulare e infine l'invecchiamento e la morte cellulare? Il vantaggio principale di questa tecnica è che possiamo tracciare le cellule madri, tracciare la loro durata di vita e i vari marcatori molecolari allo stesso tempo, in modo da poter cercare eventi molecolari correlati alla durata della vita.

Sebbene questo metodo possa fornire informazioni sull'invecchiamento del lievito, può essere applicato anche ad altre aree, come il monitoraggio delle risposte secondarie alla partecipazione che richiede il controllo cellulare per molte generazioni. In generale, le persone che non conoscono questo metodo avranno difficoltà perché la fabbricazione di dispositivi microfluidici, il caricamento delle cellule, l'installazione del microscopio richiedono molta pratica per essere eseguiti con successo. La dimostrazione preliminare di questo metodo è fondamentale in quanto è difficile caricare correttamente il dispositivo microfluidico con lievito.

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