Single Cell - Viability and Analysis - Ampha Z32 Impedance Flow Cytometry
i.e. Pollen Count and Viability Analysis
Single cell Viability Analysis by Impedance Flow Cytometry - Ampha Z32
Amphasys’ new-generation flow cytometer is a plug & play device designed to blend simplicity with flexibility. The absence of optical components minimizes set-up times and maintenance to a minimum. Time-consuming sample preparation procedures and expensive fluorescent dyes do not belong to Amphasys’ world, because it is exclusively founded on electrical measurements. Latest generation digital signal processing electronics are specifically tailored to Amphasys’ semi-disposable chips, such that best-of-class impedance analysis on single cells is achieved.
The small dimensions of Ampha Z32 and the robustness of the electrical measurement provide a mobile and portable solution for any kind of cell analysis, be it in your laboratory or out-side in the field. Designed initially for research purposes, Ampha Z32 is now particularly suitable for routine applications, where detailed, high-quality cell analyses and ease-of-use are major requirements.
The heart of the cytometer consists of a microfluidic chip equipped with microelectrodes. This high-sensitivity sensor measures changes of the electrical resistance of the fluidic medium when cells or other particles pass through the applied AC electric field.
In order to ensure highest possible sensitivity Amphasys has developed a number of chips with different channel dimensions ranging from 15 to 250 μm. The recent production of the new 250 μm chip marks a milestone in chip design and is a breakthrough for the application to large pollen. The whole range from very small cells like bacteria to very large cells such as certain pollen grains like pumpkin, maize or cotton can now be analyzed. Chips can be used for up to several hundred measurements when treated with care.
Reagents and Disposables
Amphasys provides specialized reagents and consumables for many kinds of cells and applications as well as cleaning solutions for the care of the fluidic system.
The electric signal recorded by the highly sensitive electrodes is influenced by the cell itself and by the surrounding liquid. The contents of the buffers may have an effect to the biochemical behavior of the cells as well as the conductivity of the liquid and therefore on the signal. For these reasons, the conditions of the buffers including solvents, additives, salts, pH etc. have been optimized to keep cells viable and to obtain highest measurement sensitivity at the same time.
Ampha Z32 is designed to bridge the gap between the lab and the field. Its weight of about 8 kg and dimensions of 25 x 27 x 35 cm make it perfectly suited as a table-top lab instrument as well as a mobile field instrument. With the optionally available mobility package including a 24 V rechargeable battery and a 24 V truck adapter cable it can be used independent from a power outlet.
The instrument is of a robust build and can easily be transported in the trunk or on the back seat of a car. However, we also provide a heavy duty aluminum transport casing if you plan to move the instrument on a regular basis.
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Pollen Viability Analysis
The reliable determination of pollen quality is a key success factor in the seed production process. Experiments show a clear correlation between the viability (also called vitality) of pollen used for pollination and the obtained seed yield. External factors such as light, rain, temperature or chemical agents may influence pollen viability. Amphasys provides an easy method to monitor pollen viability and thus a powerful tool to optimize the plant breeding and seed production process. This is particularly important if laborious manual steps are involved and if pollen needs to be stored or transported from one site to another. Our technology can be used on-site, directly in or near the greenhouse.
Pollen is the “seed” for the seed industry. On the basis of this vital role it should not come as a surprise that with the availability of an efficient and reproducible technique for the monitoring of pollen quality it will soon get a lot more attention.
Pollen Ploidy Analysis
Analyses performed on Ampha Z32 record impedance amplitude and phase signals of every single cell.
These are displayed on dot plots and provide size and viability information, respectively. Pollen size of haploid and diploid tomato differ significantly which can be used in order to observe ploidy.
Developmental Stages of Pollen
Multifrequency analyses in the radio frequency range interrogate electrical membrane and cytoplasmic characteristics that change during pollen maturation. It has been shown with brassica pollen, that the development from uni-nucleate to tri-cellular pollen can be followed by impedance flow cytometry. Therefore, Ampha Z32 allows to characterize the developmental stages of pollen.
|Available sensing channel dimensions||15 x 15 μm, 30 x 30 μm, 50 x 50 μm, 80 x 80 um, 120 x 120 μm or 250 x 250 μm
(depending on application requirements)
|Frequency range||0.1 MHz – 30 MHz|
|Frequency selection||Up to 4 different frequencies simultaneously|
|Sample volume||50-2000 μl (typically)|
|Concentration range||1 x 10(3) to 1 x 10(7)cells/ml|
|Particle size||1 - 60 μm (up to 150 μm with 250 μm chip)|
|Sample flow rate||5 – 2500 μl/min (depending on channel dimension)|
|Pump||Peristaltic pump with disposable pump head|
|Labware compatibility||Standard 5 ml polystyrene round - bottom tubes (Falcon® 352058)|
Operating System (min requirement)
|Laptop with Windows® 7 or 10|
|15” TFT LCD display|
|i5 or i7 QuadCore processor|
|128 GB SSD|
Dimensions & Weight
|Dimensions (W x D x H)||255 x 275 x 353 mm|
|Temperature||16° - 32°C|
|Humidity||10% - 90% relative non - condensing|
|Power||24V DC ± 10% , max. 3A, < 90 W|