Automated Live Cell Imaging

Applications

Image Tissues, Suspension Cells, and Adherent Cells with 100 Focal Planes

  • The CellAssist captures bright-field, and phase contrast images at up to 100 focal planes, 2 µm to 50 µm apart.

  • Users can select the number of focal planes, and distances between each focal plane depending on the experiment.

  • Each image is five megapixels, and 500 to 20,000 images are captured, rapidly stitched together, and added to a centralized database.

  • Gain deeper insights into cell morphology with the CellAssist!

Neurosphere differentiating into a neuron, imaged on the CellAssist in a 96-well round-bottom plate at 10x in bright-field

Quantitatively Assess Cell Growth, and Confluence

CellAssist measures cell growth through doubling time measurement:

  • CellAssist provides several options for measuring cell growth:
    • Doubling time
    • Doubling rate per day
    • Doubling rate per hour

  • Allows for manually defined starting, and ending points for accurate growth rate determination

  • Enables the user to more accurately assess confluence within the selected data set to better predict passage times

HEK293, and A549 in each well of a 6 well plate. Images were collected every 4 hours for a period of 5 days. During the course of this experiment, 27 standard scans (4x brightfield) were collected, and automatically analyzed for confluence by the CellAssist software.

Confluence Measurement in Center, and Outer Rings of the Well

Objective metrics, and images for consistent decision-making

CellAssist objectively measures confluence in the center, and more difficult to read outer ring of the well:

  • Compare, and optimize processes

  • Identify outliers that are evidence of process problems

  • Forecast optimal passaging times

Whole Cell Imaging, and Segmentation

The CellAssist offers the option to acquire quick scan images (selected areas of the well), or expertly stitched whole well images
  • The user is able to view images by spatial coordinates throughout time, enabling robust time-lapse imaging for any region of interest (including that of the whole well)​

  • A stitched whole well image of a 6-well plate is shown on the right, with colony outlines identified by Thrive Bioscience algorithms​

Deeper Insight into Cell Morphology with Time-Series Images

Daily scans of 6-well plates at 4x shows the capability of CellAssist to follow colonies or a region of interest over time
Even though cells are in standard format off-the-shelf-plates, CellAssist provides excellent image registration across multiple scans.
Day 1

Day 2

Day 3

Day 4

Analysis of Outliers to Improve Quality

CellAssist improves quality, and documentation of cells

Problem: Current tools do not collect images, and data along with tools that enable users to visualize outliers. Outliers are important because they may indicate problems in quality that require investigation.

Objective: Document, and analyze images, and data on cells over time to improve quality, and reproducibility of cell, tissue, and suspension culture.

CellAssist Solution: CellAssist provides comparable data over time that allows for identification of outliers.

(In this example, upon further investigation, the customer found cases of mislabeling.)

(Statistical anomaly highlighted)

Monoclonality Probability of Candidate Colonies

CellAssist improves quality, and increases yields by using data to grade, and pick colonies

Problem: Gene editing transfection rates can be poor, so it is important that monoclonal colonies be selected for passaging.

Objective: Accurately determine whether a colony is monoclonal by capturing series of time lapse images with excellent registration across scans.

 CellAssist Solution: The CellAssist time-lapse imaging enables the lineage of candidate colonies to be tracked to a single starting cell. 

Morphological Assays for iPSC to Glial Cell Transformations

Morphological assays do not disturb cell growth

Problem: Differentiation of neuronal progenitor cells into glial cells was measured using fluorescence-based end point assays, which did not allow for further growth, and study of the cells. An equivalent assay was needed based on imaging of live cells.

Objective: Acquire high resolution images that are comparable over time in order to study differentiation without dyes, and/or probes that interfere with the cells and colonies.

CellAssist Solution: The CellAssist’s high resolution stacks of images, with highly accurate registration across scans, enabled the characterization of cell morphology without disturbing the cells, and colonies.

Comparison of Cell Growth on Various Plate Surface Coatings

Optimization, Yield, and Cost Savings

Problem: (a) The coating of plates can profoundly affect the survivability, and growth of colonies (in this case gene edited ones starting with very small quantities). (b) Some coatings are also significantly more expensive. 

Objectives: Compare two coatings to choose the optimal one by characterizing, and comparing the cells under both conditions over time. (b) Tailor, test, and directly compare the impacts of new protocols to select the best coating, AND protocol.

CellAssist Solution: The CellAssist’s documentation, colony counting curves, and examination of images are used to characterize the number of colonies for each condition, and the results are collated, and easily compared across conditions.