EVOS Onstage Incubator

Catalog number: AMC2000

The second-generation EVOS Onstage Incubator (OSI-2) is an accessory for EVOS M5000 and EVOS M7000 Imaging Systems that enables the incubation of cells at user-defined temperature, humidity, and gas (O2, CO2 or N2), for capture of images and recording of time-lapse movies of live-cells under physiological and non-physiological conditions (e.g., hypoxia) over long periods of time.

Key features

  • Intuitive user-interface (UI) simplifies experiment set-up and monitoring of experimental conditions
  • UI enables adjustments while experiments are running
  • Small footprint and sleek design saves lab space
  • Onboard air compressor eliminates need for separate air-tank; OSI-2 is also quiet at 42 dB
  • Precise control over temperature (see figure below), humidity, and gasses provides relatively uniform live-cell environment whether physiological or non-physiological conditions (e.g., hypoxia)
     

Temperature uniformity for OSI-2 with a 37°C setpoint.

Videos and data

OSI-2 maintains the optimal live-cell environment for your live-cell experiment, including neurite outgrowth, cell proliferation, and hypoxia. Images and time-lapse movies can be further analyzed and additional visualization options are available with Celleste 6.0 Image analysis software.
 

Lysosomal localization and degradation

Time-lapse imaging over 65 hours of lysosomal localization and degradation with a LysoLight Deep Red conjugated antibody using the EVOS Onstage Incubator and EVOS M7000 microscope. SKBR3 cells were treated with 1 µg/mL of LysoLight Deep Red conjugated to the chemotherapy antibody, trastuzumab and high-resolution images were captured every 15 minutes over 3 days.

 

Cell proliferation

Time-lapse imaging of U2OS cells with an EVOS M7000 Imaging System equipped with an EVOS Onstage Incubator (OSI-2). U2OS treated with CellLight GFP-tubulin (C10613) and imaged every 15 minutes over 48 hours at 37oC, 5% CO2, and 80% humidity with an EVOS M7000 Imaging System (Cat. No. AMF7000) equipped with an OSI-2 (Cat. No. AMC2000) and a 20X objective (Cat. No. AMEP4734).

 

Neurite outgrowth

Time-lapse imaging of neurite outgrowth from neurons over several days using an EVOS M7000 Imaging System equipped with an EVOS Onstage Incubator (OSI-2). Rat hippocampal neurons plated on Poly-D-Lysine (Cat. No. A38904) coated plates in Neurobasal Plus medium (Cat. No. A3582901) with B-27 Plus supplement (Cat. No. A35828) incubated in an OSI-2 (Cat. No AMC2000) with 5% CO2, 37°C, and 80% humidity and imaged every 15 minutes for 48 hours with an EVOS M7000 Imaging System (Cat. No. AMF7000) equipped with a 20x objective (Cat. No. AMEP4734). Left image: day 1 neurite outgrowth; right image: day 3 neurite outgrowth.

 

Hypoxia


Real-time detection of hypoxia with Image-iT Hypoxia Red using an EVOS M7000 Imaging System equipped with an EVOS Onstage Incubator (OSI-2). A549 cells were labeled with Image-iT Hypoxia Red (Cat. No. H10498) and incubated in the OSI-2 (Cat. No. AMC2000) at 20% oxygen for 1 hour (left image) then oxygen was decreased to 1% oxygen for 1 hour (right image) with 5% CO2, 37oC, and 80% humidity held constant. Imaging performed with an EVOS M7000 Imaging System (Cat. No. AMF7000), equipped with a custom EVOS filter cube with GFP excitation (470/22) and RFP emission (593/40) and a 20x objective (Cat. No. AMEP4734).

Specifications

Attribute Details
Compatible vessels

Multi-well plates, 35, 60 and 100 mm petri dishes, T-25 flasks, chamber slides*
Temperature range

30°C to 40°C
CO2 range

0-20%
O2 range

0% to ambient
Humidity range 70-90% at 37-40°C
Dimensions (H X D X W)

27 x 17 x 4.1 cm (Environmental chamber)

42 x 21 x 23 cm (Control unit)
Weight

0.73 kg (Environmental chamber)

6.7 kg (Control unit)

Compatible instruments

 EVOS M5000 and EVOS M7000
*Requires appropriate vessel holder; browse EVOS vessel holders (with link to vessel holder document)

Onstage incubator vessels and vessel holders

To ensure proper operation of the EVOS Onstage Incubator, we recommend choosing vessels designed specifically for live-cell imaging. Conventional plastic plates can generate optical problems or even autofluorescence. Additionally, it is important to use only EVOS Onstage incubator compatible vessel holders and stage plates.
 

Onstage incubator media

Many cell culture media are carbonate buffered and, once removed from the incubator, the relatively lower CO2 concentration of air will cause an increase in the medium pH within an hour. The optically clear, physiological solution (Live Cell Imaging Solution, or LCIS) maintains physiological pH and osmolarity for up to 4 hours at ambient atmosphere and temperature. This is especially useful for imaging outside of an incubator but can also help to maintain pH while using the EVOS Onstage Incubator-2.

 

For longer-term imaging, particularly in a microscope-stage incubator, we offer a clear cell culture medium, Gibco FluoroBrite DMEM, that is based on DMEM to help preserve cell health while also providing low background fluorescence (comparable to that of phosphate-buffered saline and 90% lower than standard phenol red–free DMEM).

 

For Research Use Only. Not for use in diagnostic procedures.