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Beta Pack

Live Cell Imaging

Keep ideal temperature and see your cells' development in real time
Accurate temperature control

Keep your cells at 37°C ± 0.5°C outside the CO2 incubator

Up to 3 simultaneous devices

Test different conditions in the same system

For static or flow conditions

Designed for plates, dishes and perfused chips

WHAT IS A BETA PACK?

Beta Packs, such as the Live Cell Imaging pack, are an assembly of instruments that include Elveflow’s catalog products and, at least, one beta instrument, in this case, the stage top incubator. It has recently been developed by our Innovation team, and it is in testing phase. If you are interested in this beta pack, its applications, or possible variations to better fit your needs, contact our experts!

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Features & benefits

 live cell imaging under perfusion

 

Our stage top incubator allows you to perform long-term live cell imaging experiments on top of the microscope stage at ease. Designed to fit k-frame microscope stages, the internal chamber fits up to three parallel chips. The ITO glass ensures homogeneous temperature across the microfluidic devices and the frame was made so you don’t need to worry about keeping your reservoirs heated. Your media will arrive at the right temperature on top of your cells regardless of whether you leave the reservoir at room temperature.

 

how does it work?

 

As a proof-of-concept of the suitability of the stage top incubator for live cell imaging, we cultured mouse microglial cells for 48 hours under continuous perfusion on top of the microscope stage. The media flow control was done with the OB1 pressure driven flow controller and MFS flow sensor, as shown below.

Microglial cells, as other cell types from the brain parenchyma, are very sensitive to shear stress as they are not usually exposed to flow in their physiological environment. Thus, to prevent the negative effects of shear stress, we employed the µ-Slide III 3D Perfusion and a microfluidic resistance  to ensure a low and stable flow.

 

live cell imaging setup with stage top incubator, inverted microscope, pressure driven flow controller and flow sensor

Components of live cell imaging pack

– Stage top incubator

– OB1 pressure driven flow controller

– MFS flow sensor 

– u-Slide III 3D Perfusion

Details of the experimental design can be found on the Specifications tab.

Experimental Details

Primary microglial cells were isolated from mice pups brains according to the protocol published in Sepulveda et al, Glia, 2016.

 Primary mouse microglial cells were prepared and kindly provided by Patrick P. Michel and Rocio Gimenez, from the Brain and Spine Institute in the frame of the LOCAI project.

Cells were passaged after 1 week of isolation onto ibidi chips (µ-Slide III 3D Perfusion, cat. 80376) at a density of 150 000 cells/mL. Cell medium was supplemented with FBS and PS. HEPES in powder was added to a final concentration of 20 mM. Once the HEPES dissolved, the pH was corrected and brought to 7.2 using Na 1 N, then the media was filtered using a 0,22 filter. Cells were left to attach overnight in an incubator at 37°C and 5% CO2 prior to live cell imaging.

HARDWARE

  •       OB1, MK3+
  •       1 Flow sensors MFS2
  •       Tubings (1/32” ID), fittings and reservoirs
  •       IBIDI µ-Slide III 3D perfusion
  •       8 cm of 100 µm inner diameter microfluidic resistance
  •       Microfluidic male and female Luer to ¼”-28 connectors
  •       Laminar flow hood
  •      Standard Incubator for cell attachment, stage top incubator for experiment

CHEMICALS

  •     Dulbecco’s modified Eagle’s medium (DMEM)
  •     Penicillin/Streptomycin, 10,000 U/ml, 10mg/ml Streptomycin, PAN biotech
  •     FBS, 0.2 µm sterile filtered, PAN Biotech
  •     HEPES, to buffer pH in absence of CO2 control.

SOFTWARE

    •  ESI software
    • Thermal chamber software

DESIGN OF THE CHIP

Interface type Female Luer
Well diameter 5.5 mm
Well height 1.7 mm
Volume per well 30 μl
Material Ibidi Polymer Coverslip
Surface treatment ibiTreat
Growth area per well 25 mm2

For all the specifications of the stage top incubator and it’s compatibilities, please check the dedicated page here.

The objective of our Beta Packs is to gauge researchers’ interest in specific applications by utilising a range of catalog products along with new beta instruments.

Our Beta Packs are currently in the developmental phase and undergoing testing. Our goal is to eventually transform these packs into complete application products. We value your feedback, as it helps us enhance and refine our offerings for these specific applications.

For more information, advice, or support in your research, please do not hesitate to contact us at innovation@elvesys.com. We are always happy to help and support you in your research.

Our Beta Packs, such as the Live Cell Imaging pack, available under specific conditions. As these packs are still in the developmental stage, we have some eligibility criteria to ensure their optimal success rate. A consultation with our experts is necessary to evaluate your eligibility to our Beta Testing Program.

At the Microfluidics Innovation Center, we believe in tailoring our approach to suit each researcher’s unique needs. Therefore, before purchasing a Beta Pack, we encourage you to reach out to our technical team for a detailed discussion of your research objectives and requirements.

To learn more about our Beta Packs or to schedule a consultation with our experts, please contact us at innovation@elvesys.com. We look forward to collaborating with you and supporting your research endeavors.

You can order our instruments on the product section of our website.

No, the Stage Top Incubator allows gas exchange with the atmosphere.

Live cell imaging Results

 

Live cell imaging allows for the study of the dynamic aspect of cellular processes. In vitro models, however, need to be under strict environmental control. Parameters such as pH, temperature and oxygen levels are critical for robust and reliable results. Whether the experimental paradigm requires shear stress, continuous perfusion or static conditions, the need for accurate temperature regulation is the same. 

The videos below are 8h time-lapses of primary microglial cells taken immediately after stimulation with lipopolysaccharide (LPS). These videos were recorded using microglial cells seeded onto microfluidic chips and either left under static, non-perfused conditions (video 1) or connected to a pumping system for continuous perfusion along the experiment (1 µL/min, video 2).

 

Video 1.- Microglial cells seeded in a µ-Slide III 3D Perfusion chip well and maintained in the stage-top incubator on the microscope frame in static conditions for 8 hours after LPS stimulation. Cells were challenged with 10 ng/mL LPS at t = 0 h. Micrographs in phase contrast were acquired every 30 minutes for 8 hours. Image acquisition was made using an AxioVert inverted microscope using a 20x magnification objective (Zeiss).

 

Video 2.- Microglial cells seeded in a µ-Slide III 3D Perfusion chip well and maintained in the stage-top incubator with continuous perfusion at a flow rate of 1 µL/min. Cells were challenged with 10 ng/mL LPS at t = 0 h. Micrographs in phase contrast were acquired every 20 minutes for 8 hours. Image acquisition was made using an AxioVert inverted microscope using a 20x magnification objective (Zeiss).

 

In both cases, the incubator allowed for the recording of microglial cells. Micrographs taken every 20 or 30 minutes show the highly dynamic and motile nature of these cells. Importantly, microglial cells, as other cell types from the brain parenchyma, are not exposed to mechanical stresses under physiological conditions. They require, however, nutrient delivery and removal of waste metabolites for survival as in physiological conditions.

 

The stage top incubator presented here allows for flexibility in designing experiments, including when deciding to apply specific mechanical stress linked to flow. Several off-the-shelf chips with different geometries will allow to extend the design possibilities to address either physiological or pathophysiological conditions using the tissue or cell type of choice

 

CUSTOMIZE YOUR Live cell imaging PACK

Do you use well-plates instead of microfluidic chips for live cell imaging? Your microscope doesn’t have a k-frame stage? You don’t work at all with cell biology but need to keep your samples warm on top of the microscope stage? No worries! Just email us and we will see what we can do!

Elveflow Beta Products and Packs are fully customizable to fit your needs perfectly. Our microfluidic specialists and researchers will help you choose the best instruments and accessories and accompany you during the setup of the microfluidic platform until you can obtain your first experimental results.

Elveflow can provide a wide range of different reservoirs, flow sensors, bubble detectors and traps or any relevant microfluidic instruments.

Contact our experts to answer any questions about this stage top incubator for microfluidics beta pack and how it can match your specifications!

 

– Check our other BETA PRODUCTS AND PACKS for various applications –

The LOCAI and ACDC projects results helped develop this instrument pack, with funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 824060 (ACDC project), French National Research Agency (ANR) and the German Federal Ministry of Education and Research (BMBF) in the frame of the 1st German-French Joint Call for proposals on Artificial Intelligence. Primary mouse microglial cells were prepared and kindly provided by Patrick P. Michel and Rocio Gimenez, from the Brain and Spine Institute in the frame of the LOCAI project.

logo european commission    ACDC LOCAI_logo ANR_Elvesys-microfluidics-artifical intelligenceParis Brain Institute Logo

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