In the culture and observation of cells, conventional equipment places stress on valuable cell lines, risks contamination, and can lead to decreased cell viability. The BioStation CT automatically conducts operations from culture to observation of cells under optimal conditions in the same incubator, according to a user-configured schedule. Moreover, culture condition records are saved with image data for worry-free culture of important samples.

Cell viability is dependent on the performance of the CO₂ incubation system. Nikon’s BioStation employs powerful functionality for stable environmental control of CO₂, temperature, and humidity, reducing stress on cells; thereby enhancing viability.

• Temperature Management - Direct heating system that accurately maintains a constant temperature is employed for temperature management. Sheet type heaters built into the incubator’s six sides directly control the temperature.

• Humidity Management - Humidification system is employed for humidity management. The risk of contamination, common in standard incubation sytems, has been reduced through the use of an air jet.

• Reduction of Stress on Cells - An access gate is used to minimize transient environmental fluctuations when placing samples in the carrier. The access gate is a much smaller area then the entire incubator door.

• Accurate Records of Environmental Changes - The control unit is equipped with a computer that records environmental changes along the time axis. It allows secure management of three major elements (temperature, humidity and CO₂ concentration) of the incubator and saves environmental changes linked to images as a log.

1. Full Well Scan Observation

In this mode, BioStation CT, with its advanced stage and intuitive software allows researchers to automate the process of imaging the entire area of a well or all wells within a 6, 12, 24, 48, or 96 well-plate format. Simply select “full well scan” when defining the automated time-lapse schedule and the BioStation CT will calculate the amount of frames necessary to complete the reconstructed well area.

Left image: 6-well plate shown taken by the macro color camera.

Right image: full well-scans in phase reconstructed to reveal the entire area of each well.

Full well scans of developing iPS colonies grown in the BioStation CT and periodically removed for media exchange. Images are zoomed so that colonies can be seen without loss of resolution.  

Images courtesy of Konrad Hochedlinger, Harvard Stem Cell Institute.

2. Multi-Vessel Observation

In this mode, BioStation CT shuttles each vessel programmed for automated time-lapse imaging between the culture vessel stocker (30 plate positions) and the microscope stage for acquisition. Once the imaging cycle is complete the plate is returned to its stocker position and the next plate is selected to be brought to the stage.

3. High-Speed Observation

In this mode, BioStation CT selects the vessel from the stocker and transports it to the microscope stage where it stays for the duration of the entire imaging cycle for all acquisitions. Higher temporal resolution is possible by alleviating the need to shuttle the plate back and forth from the stocker to the stage for each acquisition.

The enclosed inverted microscope offers sophisticated imaging capabilities:

• Images captured with 1 megapixel cooled CCD camera
• Range of magnification from 2x to 40x that can be user selected depending on application requirements.
• An image acquisition system with auto focus allows simple and accurate observation of cells and storage of images.
• A newly developed observation stage — specifically designed to perform in environments with humidity of over 95% — provides a broad 120 x 86mm observation area.
• Nikon’s exclusive Apodized Phase Contrast (APC) objective lens reduces halo and provides clear images of minute intracellular structures.
• An LED illumination system that is less phototoxic to cells than conventional equipment is used for fluorescence observation.
• The phase contrast illumination, which is typically difficult to adjust, provides optimal illumination at every magnification.

• Images captured with Color CCD Camera
• Confirm handwritten vessel notes, change in culture medium color, mold generation, etc.
• Perform colorimetric analysis on pH levels within the media

The BioStation CT's unique design includes a 30 space rack that can accommodate a wide variety of vessels, such as culture flasks, 35mm dishes and multi-well plates. As 30–24-well plates can be placed in the BioStation CT, there are endless configurations for simultaneous experimentation.

The transfer unit automatically transfers culture vessels from the rack to the inverted microscope according to a configured schedule. This transfer is executed with great care to avoid media splashing and mechanical vibration.

The BioStation CT is not only for culture and observation of cells, as it places great importance on cell traceability for each experiment. Post-culture experiments can also be conducted with high reliability.

• Removing culture vessels, viewing records and preparing schedules requires the user to log in. Use of a log-in menu allows secure storage of important cells and verification of all access and manipulation.

• With an easy-to-use and visually intuitive software menu, the number of culture vessels stored in each storage space is visible at a glance. Selected vessels can be individually moved to the access gate by the transfer unit, leaving all other trays untouched.
• Acquired cell culture data (temperature, humidity, CO₂ gas concentration), name of handler, and saved images can be viewed. Remote observation from outside the laboratory via a network is also possible.
• 2 TB of on-board hard disk space for image storage.

Fluorescence

• 3-channel LED based excitation
• Preconfigured for common fluorescent dyes: GFP, CFP, DsRed
• On-the-fly phase/fluorescent overlay for co-localization studies
• For use with off-the-shelf dichroic filters
• Captures maximum intensities without losing resolution

Mixed Gas Environment

• Controls levels of CO2, O2, N2
• Mimics hypoxic environment
• Grow embryos in optimal condition
• Supports regenerative medicine applications

Image Analysis Software

• Flexible platform for the teaching of a broad range of microscopy image recognition applications
• Recipes provide a seamless and complete application execution with a single click on time-lapse movies
• Rapid and low cost application development
• Predictable performance and fast prototyping

BioStation CL-Quant is an innovative, next generation live-cell image analysis program that enables scientists to automatically detect, segment, measure, classify, analyze and discover cellular and subcellular phenotypes in their live-cell imaging experiments; in both brightfield (phase) and three-channel fluorescence. CL-Quant is a flexible platform which lets researchers ‘teach’ the computer using advanced decision-based methods for high volume execution and a broad range of microscopy image recognition applications. Pre-configured recipes are also available which provide a seamless and complete application execution with a single click on time-lapse movies.

Assay Design

To count cells, CL-Quant segments each cell individually from a phase contrast image.

Blue cells are cells with a characteristic shape which our universal cell detection recipe can measure. Here we identify that the culture is indeed growing over time by determining the distinct cell number as seen in the graph on the right.

Assay Design

To measure cell coverage, CL-Quant uses a universal recipe which can be applied to any adherent cell type.

The blue mask seen here indicates the area occupied by the cells. CL-Quant measures the confluency rates over time.

Assay Design

To measure the rate of cell division, CL-Quant compares dividing cells with the adherent cells from a brightfield image.

The red mask seen here indicates the cells undergoing mitosis. CL-Quant measures the confluency rates over time and simultaneously measures the mitotic index within this population.

Assay Design

To measure and compare multiple parameters, CL-Quant applies relevant overlapping masks.

The blue mask represents the total adherent cell population. The green mask counts the adherent cell population. The red mask counts the dividing cell population. CL-Quant allows for various ways to interpret the data by utilizing multiple masks in this way.