A Method for Determining the Weight of an Individual Yeast Cell

Haddad, Samir A.; Lindegren, Carl C.

doi:10.1128/am.1.3.153-156.1953

Cited by 18 publications

(5 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The effectiveness of the overall weighting procedure was thus validated on a biological sample: a population of yeast cells. Besides being biologically relevant, measuring the mass of single yeasts in a population can also have biotechnology relevance, for example to identify the state of the population in relation to the production of recombinant proteins [41], and it has been addressed for more than half a century [42]. Single yeast (Saccharomyces cerevisiae) cells are a few μm in size and they typically weigh 10-100pg [43], perfectly fitting the measurement capabilities of the proposed FluidFM-based approach.…”

Section: Measuring Masses From Frequency Shiftmentioning

confidence: 89%

Serial weighting of micro-objects with resonant microchanneled cantilevers

et al. 2016

View full text Add to dashboard Cite

Atomic force microscopy (AFM) cantilevers have proven to be very effective mass sensors. The attachment of a small mass to a vibrating cantilever produces a resonance frequency shift that can be monitored, providing the ability to measure mass changes down to a few molecules resolution. Nevertheless, the lack of a practical method to handle the catch and release process required for dynamic weighting of microobjects strongly hindered the application of the technology beyond proof of concept measurements. Here, a method is proposed in which FluidFM hollow cantilevers are exploited to overcome the standard limitations of AFM-based mass sensors, providing high throughput single object weighting with picogram accuracy. The extension of the dynamic models of AFM cantilevers to hollow cantilevers was discussed and the effectiveness of mass weighting in air was validated on test samples.

show abstract

Section: Measuring Masses From Frequency Shiftmentioning

confidence: 89%

Serial weighting of micro-objects with resonant microchanneled cantilevers

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Second, the wet cell mass was calculated to be 88 pg based on a specific gravity of 1.1 commonly reported for yeast. [ 55,56 ] Third, the dry cell mass was determined to obtain m cell = 22 pg using the approximation that dry matter comprises about a quarter of the wet cell mass. [ 57,58 ]…”

Section: Methodsmentioning

confidence: 99%

Fed‐Batch Droplet Nanobioreactor for Controlled Growth of Cyberlindnera (Pichia) jadinii: A Proof‐Of‐Concept Demonstration

Totlani

Wang

Bisschops

et al. 2021

Adv Materials Technologies

View full text Add to dashboard Cite

Improvement of the microorganism's product yield on feedstock is possible by targeted genetic modification, which requires a solid understanding of the species metabolism and genetics. Alternatively, genetic modifications can be introduced randomly such that the chance that a resulting mutant performs better than the original one is small. Consequently, many mutants need to be created and subsequently screened in order to identify the best performing ones. [1] Given the large number of experiments, it is common practice to grow and study all the different mutants in individual microliter-sized wells on microtiter plates. While this allows for parallel screening in an automated fashion using pipetting robots and plate readers, conventional microtiter plates lack the ability to feed nutrients and control pH by base or acid addition. It is indeed challenging to feed liquids at flow rates in the nanoliter per hour range to all the individual wells of a microtiter plate and to do so accurately. Therefore, screening of mutants is commonly performed under batch conditions with all feedstock present from the start and no further control over feedstock concentration and pH. In contrast, more than 80% of the processes in industrial biotechnology are operated under so called fed-batch conditions with control over feedstock concentration and pH. [2,3] This incompatibility between the physiological conditions during screening and industrial operation not only leads to selection of false positives which fail to generate competitive yields at industrial scale, but also fails to identify the best mutants for fed-batch conditions. [4][5][6] The effectiveness of screening hence greatly benefits from a technology that enables growing and studying a large number of microorganisms under precisely controlled conditions representative of industrial bioreactors. Besides screening of mutants, optimization of process conditions, a second important aspect in bioprocess development, also benefits from such a technology. [7] Although there has been progress in recent years, there remains both need and opportunity to cost-effectively and with fidelity miniaturize fermentation to screen under fed-batch conditions.Several strategies have been developed to study microorganisms under controlled growth conditions. One strategy is to A key bottleneck in bioprocess development is that state-of-the-art tools used for screening of cells and optimization of cultivation conditions do not represent the conditions enforced at industrial scale. At industrial scale, cell growth is strictly controlled ("fed-batch") to optimize the metabolites produced by the cells. In contrast, cell growth is uncontrolled ("batch") in microwells commonly used for bioprocess development due to the difficulty to continuously supply minute amounts of nutrients to the cells in these wells over the course of the cultivation experiment. This work addresses this bottleneck through the development of a droplet-based fed-batch nanobioreactor.A key challenge addressed in this work is ...

show abstract

“…A hundred particles were released at the inlet, with a diameter of 4 µm which corresponded to the size of a yeast cell, and 1 cell was positioned in the microwell. The density of the particles was set at 1117 kg/m 3 , which is the average value for yeast cells found in the literature [44][45][46][47].…”

Section: Computational Simulation Of Fluid Flow In the Microfluidic Chipmentioning

confidence: 99%

Single-Cell Optical Nanomotion of Candida albicans in Microwells for Rapid Antifungal Susceptibility Testing

et al. 2023

View full text Add to dashboard Cite

Candida albicans is an emerging multidrug-resistant opportunistic pathogen representing an important source of invasive disease in humans and generating high healthcare costs worldwide. The development of a rapid and simple antifungal susceptibility test (AFST) could limit the spread of this disease, increase the efficiency of treatment, and lower the risk of developing resistant strains. We developed a microfluidic chip containing an array of microwells that were designed to trap the cells and perform rapid antifungal susceptibility tests using optical nanomotion detection (ONMD). Yeast cell entrapment in a microwell allows for a very rapid exchange of growth medium with the antifungal, which enables performing single-cell ONMD measurements on the same cell before and after antifungal treatment. The exposure to a low concentration of the antifungal caspofungin or flucanozole induced a significant decrease in the nanomotion signal, demonstrating the high sensitivity of this technique. We used this chip to quantify the real-time response of individual C. albicans cells to the antifungal treatment in as fast as 10 min. This simple and label-free technique could be further developed into a simple-to-use device that allows the performance of fast AFST as part of a routine hospital procedure in developed and also eventually developing world countries.

show abstract

A Method for Determining the Weight of an Individual Yeast Cell

Cited by 18 publications

References 0 publications

Serial weighting of micro-objects with resonant microchanneled cantilevers

Serial weighting of micro-objects with resonant microchanneled cantilevers

Fed‐Batch Droplet Nanobioreactor for Controlled Growth of Cyberlindnera (Pichia) jadinii: A Proof‐Of‐Concept Demonstration

Single-Cell Optical Nanomotion of Candida albicans in Microwells for Rapid Antifungal Susceptibility Testing

Contact Info

Product

Resources

About